Difference between revisions of "User:Neil J. Parikh/Lab Notebook"

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h1
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margin-bottom:6.0pt;
 
margin-bottom:6.0pt;
 
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mso-bidi-font-family:"Times New Roman";
 
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h2
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mso-style-qformat:yes;
 
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mso-bidi-font-family:"Times New Roman";
 
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p.MsoHeader, li.MsoHeader, div.MsoHeader
 
p.MsoHeader, li.MsoHeader, div.MsoHeader
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margin-top:0in;
 
 
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margin-bottom:6.0pt;
 
margin-bottom:6.0pt;
 
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font-family:"Arial","sans-serif";
 
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font-weight:bold;
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mso-bidi-font-style:normal;}
 
 
p.MsoFooter, li.MsoFooter, div.MsoFooter
 
p.MsoFooter, li.MsoFooter, div.MsoFooter
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mso-style-parent:"Body Text";
 
margin-top:0in;
 
 
margin-right:0in;
 
margin-right:0in;
 
margin-bottom:6.0pt;
 
margin-bottom:6.0pt;
 
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font-family:"Arial","sans-serif";
 
font-family:"Arial","sans-serif";
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mso-bidi-font-family:"Times New Roman";
 
 
font-weight:bold;
 
font-weight:bold;
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font-style:italic;
 
mso-bidi-font-style:normal;}
 
 
p.MsoListBullet, li.MsoListBullet, div.MsoListBullet
 
p.MsoListBullet, li.MsoListBullet, div.MsoListBullet
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margin-top:6.0pt;
 
 
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font-size:10.0pt;
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font-family:"Arial","sans-serif";
 
mso-fareast-font-family:"Times New Roman";
 
mso-bidi-font-family:"Times New Roman";}
 
 
p.MsoListNumber, li.MsoListNumber, div.MsoListNumber
 
p.MsoListNumber, li.MsoListNumber, div.MsoListNumber
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margin-right:0in;
 
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p.MsoListBullet2, li.MsoListBullet2, div.MsoListBullet2
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margin-top:0in;
 
 
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p.MsoTitle, li.MsoTitle, div.MsoTitle
{mso-style-update:auto;
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margin-bottom:3.0pt;
 
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font-family:"Arial","sans-serif";
 
font-family:"Arial","sans-serif";
mso-fareast-font-family:"Times New Roman";
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mso-bidi-font-family:"Times New Roman";
 
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p.MsoBodyText, li.MsoBodyText, div.MsoBodyText
 
p.MsoBodyText, li.MsoBodyText, div.MsoBodyText
{mso-style-unhide:no;
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margin-top:0in;
 
 
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margin-right:0in;
 
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p.MsoSubtitle, li.MsoSubtitle, div.MsoSubtitle
 
p.MsoSubtitle, li.MsoSubtitle, div.MsoSubtitle
{mso-style-priority:11;
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mso-style-qformat:yes;
 
mso-style-link:"Subtitle Char";
 
mso-style-next:Normal;
 
 
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margin-right:0in;
Line 583: Line 119:
 
margin-left:0in;
 
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font-size:12.0pt;
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mso-fareast-font-family:"Times New Roman";
 
mso-bidi-font-family:"Times New Roman";}
 
 
a:link, span.MsoHyperlink
 
a:link, span.MsoHyperlink
{mso-style-unhide:no;
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{color:blue;
color:blue;
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text-decoration:underline;
 
text-underline:single;}
 
 
a:visited, span.MsoHyperlinkFollowed
 
a:visited, span.MsoHyperlinkFollowed
{mso-style-noshow:yes;
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{color:purple;
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mso-themecolor:followedhyperlink;
 
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p.MsoAcetate, li.MsoAcetate, div.MsoAcetate
 
p.MsoAcetate, li.MsoAcetate, div.MsoAcetate
{mso-style-noshow:yes;
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margin:0in;
 
 
margin-bottom:.0001pt;
 
margin-bottom:.0001pt;
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p.MsoListParagraph, li.MsoListParagraph, div.MsoListParagraph
 
p.MsoListParagraph, li.MsoListParagraph, div.MsoListParagraph
{mso-style-priority:34;
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mso-style-qformat:yes;
 
margin-top:0in;
 
 
margin-right:0in;
 
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margin-bottom:.0001pt;
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mso-fareast-font-family:"Times New Roman";
 
mso-bidi-font-family:"Times New Roman";}
 
 
span.MsoIntenseEmphasis
 
span.MsoIntenseEmphasis
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mso-style-qformat:yes;
 
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font-weight:bold;
 
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p.ExperimentNumber, li.ExperimentNumber, div.ExperimentNumber
 
p.ExperimentNumber, li.ExperimentNumber, div.ExperimentNumber
 
{mso-style-name:"Experiment Number";
 
{mso-style-name:"Experiment Number";
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mso-style-unhide:no;
 
mso-style-parent:Title;
 
 
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margin-right:0in;
Line 643: Line 151:
 
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font-weight:bold;
 
font-weight:bold;
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font-style:italic;
 
mso-bidi-font-style:normal;}
 
 
p.BodyTextFirst, li.BodyTextFirst, div.BodyTextFirst
 
p.BodyTextFirst, li.BodyTextFirst, div.BodyTextFirst
 
{mso-style-name:"Body Text First";
 
{mso-style-name:"Body Text First";
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mso-fareast-font-family:"Times New Roman";
 
mso-bidi-font-family:"Times New Roman";}
 
 
p.MaterialsListFirst, li.MaterialsListFirst, div.MaterialsListFirst
 
p.MaterialsListFirst, li.MaterialsListFirst, div.MaterialsListFirst
 
{mso-style-name:"Materials List First";
 
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mso-style-parent:"List Bullet";
 
 
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margin-right:0in;
Line 680: Line 171:
 
margin-bottom:.0001pt;
 
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text-indent:-.25in;
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font-family:"Arial","sans-serif";
 
font-family:"Arial","sans-serif";
mso-fareast-font-family:"Times New Roman";
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mso-bidi-font-family:"Times New Roman";
 
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mso-bidi-font-weight:normal;}
 
 
p.TableCells, li.TableCells, div.TableCells
 
p.TableCells, li.TableCells, div.TableCells
 
{mso-style-name:"Table Cells";
 
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mso-style-parent:"Body Text";
 
 
margin:0in;
 
margin:0in;
 
margin-bottom:.0001pt;
 
margin-bottom:.0001pt;
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mso-fareast-font-family:Times;
 
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p.TableHeading, li.TableHeading, div.TableHeading
 
p.TableHeading, li.TableHeading, div.TableHeading
 
{mso-style-name:"Table Heading";
 
{mso-style-name:"Table Heading";
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mso-style-parent:"Body Text";
 
 
margin-top:4.0pt;
 
margin-top:4.0pt;
 
margin-right:0in;
 
margin-right:0in;
 
margin-bottom:4.0pt;
 
margin-bottom:4.0pt;
 
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margin-left:0in;
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font-family:"Arial","sans-serif";
 
font-family:"Arial","sans-serif";
mso-fareast-font-family:"Times New Roman";
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mso-bidi-font-family:"Times New Roman";
 
font-weight:bold;
 
mso-bidi-font-weight:normal;}
 
 
p.TableBody, li.TableBody, div.TableBody
 
p.TableBody, li.TableBody, div.TableBody
 
{mso-style-name:"Table Body";
 
{mso-style-name:"Table Body";
mso-style-unhide:no;
 
mso-style-parent:"Body Text";
 
 
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margin-bottom:2.0pt;
 
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font-family:"Arial","sans-serif";
 
mso-fareast-font-family:"Times New Roman";
 
mso-bidi-font-family:"Times New Roman";}
 
 
span.SubtitleChar
 
span.SubtitleChar
 
{mso-style-name:"Subtitle Char";
 
{mso-style-name:"Subtitle Char";
mso-style-priority:11;
 
mso-style-unhide:no;
 
mso-style-locked:yes;
 
 
mso-style-link:Subtitle;
 
mso-style-link:Subtitle;
mso-ansi-font-size:12.0pt;
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mso-bidi-font-size:12.0pt;
 
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mso-ascii-font-family:Cambria;
 
mso-fareast-font-family:"Times New Roman";
 
mso-hansi-font-family:Cambria;
 
mso-bidi-font-family:"Times New Roman";}
 
 
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mso-footnote-continuation-separator:url("iGEM%20Lab%20Notebook%20Neil%20Parikh_files/header.htm") fcs;
 
mso-endnote-separator:url("iGEM%20Lab%20Notebook%20Neil%20Parikh_files/header.htm") es;
 
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mso-footer-margin:.5in;
 
mso-header:url("iGEM%20Lab%20Notebook%20Neil%20Parikh_files/header.htm") h1;
 
mso-footer:url("iGEM%20Lab%20Notebook%20Neil%20Parikh_files/header.htm") f1;
 
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div.Section1
 
div.Section1
 
{page:Section1;}
 
{page:Section1;}
 
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mso-level-tab-stop:3.0in;
 
mso-level-number-position:left;
 
margin-left:3.25in;
 
text-indent:-.25in;
 
font-family:"Courier New";
 
mso-bidi-font-family:"Times New Roman";}
 
@list l0:level8
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0A7;
 
mso-level-tab-stop:3.5in;
 
mso-level-number-position:left;
 
margin-left:3.75in;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
@list l0:level9
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0FA;
 
mso-level-tab-stop:4.0in;
 
mso-level-number-position:left;
 
margin-left:4.25in;
 
text-indent:-.25in;
 
font-family:Wingdings;}
 
@list l1
 
{mso-list-id:-132;
 
mso-list-type:simple;
 
mso-list-template-ids:-970966950;}
 
@list l1:level1
 
{mso-level-tab-stop:1.25in;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l2
 
{mso-list-id:-131;
 
mso-list-type:simple;
 
mso-list-template-ids:-1828278010;}
 
@list l2:level1
 
{mso-level-tab-stop:1.0in;
 
mso-level-number-position:left;
 
margin-left:1.0in;
 
text-indent:-.25in;}
 
@list l3
 
{mso-list-id:-130;
 
mso-list-type:simple;
 
mso-list-template-ids:-398419408;}
 
@list l3:level1
 
{mso-level-tab-stop:.75in;
 
mso-level-number-position:left;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l4
 
{mso-list-id:-129;
 
mso-list-type:simple;
 
mso-list-template-ids:-1763429746;}
 
@list l4:level1
 
{mso-level-tab-stop:.5in;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l5
 
{mso-list-id:-128;
 
mso-list-type:simple;
 
mso-list-template-ids:-1884680332;}
 
@list l5:level1
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:1.25in;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l6
 
{mso-list-id:-127;
 
mso-list-type:simple;
 
mso-list-template-ids:-233927482;}
 
@list l6:level1
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:1.0in;
 
mso-level-number-position:left;
 
margin-left:1.0in;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l7
 
{mso-list-id:-126;
 
mso-list-type:simple;
 
mso-list-template-ids:520379406;}
 
@list l7:level1
 
{mso-level-number-format:bullet;
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:.75in;
 
mso-level-number-position:left;
 
margin-left:.75in;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l8
 
{mso-list-id:-125;
 
mso-list-type:simple;
 
mso-list-template-ids:-1918614362;}
 
@list l8:level1
 
{mso-level-number-format:bullet;
 
mso-level-style-link:"List Bullet 2";
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:.5in;
 
mso-level-number-position:left;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l9
 
{mso-list-id:-119;
 
mso-list-type:simple;
 
mso-list-template-ids:1095369742;}
 
@list l9:level1
 
{mso-level-number-format:bullet;
 
mso-level-style-link:"List Bullet";
 
mso-level-text:\F0B7;
 
mso-level-tab-stop:.25in;
 
mso-level-number-position:left;
 
margin-left:.25in;
 
text-indent:-.25in;
 
font-family:Symbol;}
 
@list l10
 
{mso-list-id:89861118;
 
mso-list-template-ids:1941491386;}
 
@list l10:level1
 
{mso-level-tab-stop:0in;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l10:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:.5in;
 
mso-level-number-position:left;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l10:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:.75in;
 
mso-level-number-position:left;
 
margin-left:.1in;
 
text-indent:.65in;}
 
@list l10:level4
 
{mso-level-text:"\(%4\)";
 
mso-level-tab-stop:1.0in;
 
mso-level-number-position:left;
 
margin-left:1.0in;
 
text-indent:-.25in;}
 
@list l10:level5
 
{mso-level-number-format:alpha-lower;
 
mso-level-text:"\(%5\)";
 
mso-level-tab-stop:1.25in;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l10:level6
 
{mso-level-number-format:roman-lower;
 
mso-level-text:"\(%6\)";
 
mso-level-tab-stop:1.5in;
 
mso-level-number-position:left;
 
margin-left:1.5in;
 
text-indent:-.25in;}
 
@list l10:level7
 
{mso-level-tab-stop:1.75in;
 
mso-level-number-position:left;
 
margin-left:1.75in;
 
text-indent:-.25in;}
 
@list l10:level8
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:2.0in;
 
mso-level-number-position:left;
 
margin-left:2.0in;
 
text-indent:-.25in;}
 
@list l10:level9
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:2.25in;
 
mso-level-number-position:left;
 
margin-left:2.25in;
 
text-indent:-.25in;}
 
@list l11
 
{mso-list-id:287706635;
 
mso-list-type:hybrid;
 
mso-list-template-ids:1137848646 -2109568246 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l11:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:1.25in;
 
text-indent:-.25in;
 
mso-ansi-font-style:normal;}
 
@list l11:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
margin-left:1.75in;
 
text-indent:-.25in;}
 
@list l12
 
{mso-list-id:413170209;
 
mso-list-type:hybrid;
 
mso-list-template-ids:786721046 67698707 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l12:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l13
 
{mso-list-id:508057793;
 
mso-list-template-ids:-259592024;}
 
@list l13:level1
 
{mso-level-tab-stop:0in;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l13:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:.5in;
 
mso-level-number-position:left;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l13:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:.75in;
 
mso-level-number-position:left;
 
margin-left:.1in;
 
text-indent:.65in;}
 
@list l13:level4
 
{mso-level-text:"\(%4\)";
 
mso-level-tab-stop:1.0in;
 
mso-level-number-position:left;
 
margin-left:1.0in;
 
text-indent:-.25in;}
 
@list l13:level5
 
{mso-level-number-format:alpha-lower;
 
mso-level-text:"\(%5\)";
 
mso-level-tab-stop:1.25in;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l13:level6
 
{mso-level-number-format:roman-lower;
 
mso-level-text:"\(%6\)";
 
mso-level-tab-stop:1.5in;
 
mso-level-number-position:left;
 
margin-left:1.5in;
 
text-indent:-.25in;}
 
@list l13:level7
 
{mso-level-tab-stop:1.75in;
 
mso-level-number-position:left;
 
margin-left:1.75in;
 
text-indent:-.25in;}
 
@list l13:level8
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:2.0in;
 
mso-level-number-position:left;
 
margin-left:2.0in;
 
text-indent:-.25in;}
 
@list l13:level9
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:2.25in;
 
mso-level-number-position:left;
 
margin-left:2.25in;
 
text-indent:-.25in;}
 
@list l14
 
{mso-list-id:513572839;
 
mso-list-type:hybrid;
 
mso-list-template-ids:1063057342 67698713 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l14:level1
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l15
 
{mso-list-id:654064786;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-352949782 67698707 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l15:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l16
 
{mso-list-id:661544068;
 
mso-list-type:hybrid;
 
mso-list-template-ids:893798754 67698707 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l16:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l16:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l16:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:1.75in;
 
text-indent:-9.0pt;}
 
@list l17
 
{mso-list-id:725564168;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-713246640 67698703 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l17:level1
 
{mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l18
 
{mso-list-id:780953595;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-1008036830 67698713 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l18:level1
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l19
 
{mso-list-id:875889331;
 
mso-list-template-ids:1480984720;}
 
@list l19:level1
 
{mso-level-style-link:"List Number";
 
mso-level-tab-stop:0in;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l19:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:.5in;
 
mso-level-number-position:left;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l19:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:.75in;
 
mso-level-number-position:left;
 
margin-left:1.0in;
 
text-indent:-.25in;}
 
@list l19:level4
 
{mso-level-text:"\(%4\)";
 
mso-level-tab-stop:1.0in;
 
mso-level-number-position:left;
 
margin-left:1.0in;
 
text-indent:-.25in;}
 
@list l19:level5
 
{mso-level-number-format:alpha-lower;
 
mso-level-text:"\(%5\)";
 
mso-level-tab-stop:1.25in;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l19:level6
 
{mso-level-number-format:roman-lower;
 
mso-level-text:"\(%6\)";
 
mso-level-tab-stop:1.5in;
 
mso-level-number-position:left;
 
margin-left:1.5in;
 
text-indent:-.25in;}
 
@list l19:level7
 
{mso-level-tab-stop:1.75in;
 
mso-level-number-position:left;
 
margin-left:1.75in;
 
text-indent:-.25in;}
 
@list l19:level8
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:2.0in;
 
mso-level-number-position:left;
 
margin-left:2.0in;
 
text-indent:-.25in;}
 
@list l19:level9
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:2.25in;
 
mso-level-number-position:left;
 
margin-left:2.25in;
 
text-indent:-.25in;}
 
@list l20
 
{mso-list-id:1327439790;
 
mso-list-type:hybrid;
 
mso-list-template-ids:115351768 67698707 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l20:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:.25in;
 
text-indent:-.25in;}
 
@list l21
 
{mso-list-id:1438332439;
 
mso-list-type:hybrid;
 
mso-list-template-ids:2107017872 67698707 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l21:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:1.0in;
 
text-indent:-.25in;}
 
@list l22
 
{mso-list-id:1718239408;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-1271611110 67698715 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l22:level1
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
text-indent:-.25in;}
 
@list l23
 
{mso-list-id:1720205758;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-1195606094 67698713 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l23:level1
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l24
 
{mso-list-id:1895461054;
 
mso-list-type:hybrid;
 
mso-list-template-ids:2059586448 67698707 67698713 67698715 67698703 67698713 67698715 67698703 67698713 67698715;}
 
@list l24:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l24:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l24:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:1.75in;
 
text-indent:-9.0pt;}
 
@list l25
 
{mso-list-id:2097238860;
 
mso-list-template-ids:-266300996;}
 
@list l25:level1
 
{mso-level-tab-stop:.25in;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l25:level2
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:.5in;
 
mso-level-number-position:left;
 
margin-left:.75in;
 
text-indent:-.25in;}
 
@list l25:level3
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:.75in;
 
mso-level-number-position:left;
 
margin-left:.1in;
 
text-indent:.65in;}
 
@list l25:level4
 
{mso-level-text:"\(%4\)";
 
mso-level-tab-stop:1.0in;
 
mso-level-number-position:left;
 
margin-left:1.0in;
 
text-indent:-.25in;}
 
@list l25:level5
 
{mso-level-number-format:alpha-lower;
 
mso-level-text:"\(%5\)";
 
mso-level-tab-stop:1.25in;
 
mso-level-number-position:left;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l25:level6
 
{mso-level-number-format:roman-lower;
 
mso-level-text:"\(%6\)";
 
mso-level-tab-stop:1.5in;
 
mso-level-number-position:left;
 
margin-left:1.5in;
 
text-indent:-.25in;}
 
@list l25:level7
 
{mso-level-tab-stop:1.75in;
 
mso-level-number-position:left;
 
margin-left:1.75in;
 
text-indent:-.25in;}
 
@list l25:level8
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:2.0in;
 
mso-level-number-position:left;
 
margin-left:2.0in;
 
text-indent:-.25in;}
 
@list l25:level9
 
{mso-level-number-format:roman-lower;
 
mso-level-tab-stop:2.25in;
 
mso-level-number-position:left;
 
margin-left:2.25in;
 
text-indent:-.25in;}
 
@list l26
 
{mso-list-id:2119710673;
 
mso-list-type:hybrid;
 
mso-list-template-ids:-968488302 -2109568246 67698707 67698713 67698713 67698713 67698715 67698703 67698713 67698715;}
 
@list l26:level1
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:1.25in;
 
text-indent:-.25in;
 
mso-ansi-font-style:normal;}
 
@list l26:level2
 
{mso-level-number-format:roman-upper;
 
mso-level-tab-stop:none;
 
mso-level-number-position:right;
 
margin-left:1.25in;
 
text-indent:-.25in;}
 
@list l26:level3
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-9.0pt;}
 
@list l26:level4
 
{mso-level-number-format:alpha-lower;
 
mso-level-tab-stop:none;
 
mso-level-number-position:left;
 
text-indent:-.25in;}
 
@list l19:level2 lfo13
 
{mso-level-start-at:-1073762832;}
 
@list l19:level3 lfo13
 
{mso-level-start-at:38108040;}
 
@list l19:level4 lfo13
 
{mso-level-start-at:-1073762928;}
 
@list l19:level5 lfo13
 
{mso-level-start-at:37978784;}
 
@list l19:level6 lfo13
 
{mso-level-start-at:50823780;}
 
@list l19:level7 lfo13
 
{mso-level-start-at:37970320;}
 
@list l19:level8 lfo13
 
{mso-level-start-at:268374015;}
 
@list l19:level9 lfo13
 
{mso-level-start-at:806748160;}
 
ol
 
 
{margin-bottom:0in;}
 
{margin-bottom:0in;}
 
ul
 
ul
 
{margin-bottom:0in;}
 
{margin-bottom:0in;}
.style1 {color: #FFFFFF}
 
 
-->
 
-->
 
</style>
 
</style>
<!--[if gte mso 10]>
+
 
<style>
 
/* Style Definitions */
 
table.MsoNormalTable
 
{mso-style-name:"Table Normal";
 
mso-tstyle-rowband-size:0;
 
mso-tstyle-colband-size:0;
 
mso-style-noshow:yes;
 
mso-style-priority:99;
 
mso-style-qformat:yes;
 
mso-style-parent:"";
 
mso-padding-alt:0in 5.4pt 0in 5.4pt;
 
mso-para-margin:0in;
 
mso-para-margin-bottom:.0001pt;
 
mso-pagination:widow-orphan;
 
font-size:10.0pt;
 
font-family:"Times New Roman","serif";}
 
table.MsoTableGrid
 
{mso-style-name:"Table Grid";
 
mso-tstyle-rowband-size:0;
 
mso-tstyle-colband-size:0;
 
mso-style-unhide:no;
 
border:solid windowtext 1.0pt;
 
mso-border-alt:solid windowtext .5pt;
 
mso-padding-alt:0in 5.4pt 0in 5.4pt;
 
mso-border-insideh:.5pt solid windowtext;
 
mso-border-insidev:.5pt solid windowtext;
 
mso-para-margin:0in;
 
mso-para-margin-bottom:.0001pt;
 
mso-pagination:widow-orphan;
 
font-size:10.0pt;
 
font-family:"Times New Roman","serif";}
 
</style>
 
<![endif]--><!--[if gte mso 9]><xml>
 
<o:shapedefaults v:ext="edit" spidmax="4098"/>
 
</xml><![endif]--><!--[if gte mso 9]><xml>
 
<o:shapelayout v:ext="edit">
 
  <o:idmap v:ext="edit" data="1"/>
 
</o:shapelayout></xml><![endif]-->
 
 
</head>
 
</head>
  
<body lang=EN-US link=blue vlink=purple style='tab-interval:.5in'>
+
<body lang=EN-US link=blue vlink=purple>
  
 
<div class=Section1>
 
<div class=Section1>
  
 
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
 
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
  style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt:
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  style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
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border:none'>
  <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
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  <tr>
 
   <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 
   <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
  background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto;
 
 
   padding:0in 5.4pt 0in 5.4pt'>
 
   padding:0in 5.4pt 0in 5.4pt'>
   <p class=MsoTitle style1 style='margin-right:26.1pt'>Pipetting &amp; Making
+
   <p class=MsoTitle style='margin-right:26.1pt'>Pipetting &amp; Making
   Solutions</p> </td>
+
   Solutions</p>
 +
  </td>
 
   <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 
   <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
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   padding:0in 5.4pt 0in 5.4pt'>
 
   padding:0in 5.4pt 0in 5.4pt'>
   <p class=ExperimentNumber style1>Pg. 6</p> </td>
+
   <p class=ExperimentNumber>Pg. 6</p>
 +
  </td>
 
  </tr>
 
  </tr>
 
</table>
 
</table>
  
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span
-.25in;mso-list:l24 level1 lfo21'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span
+
</span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Pipetting<o:p></o:p></span></u></span></p>
+
font-weight:normal;font-style:normal'>Pipetting</span></u></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Learned to use pipettes properly</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>Learned to use
 
pipettes properly<u><o:p></o:p></u></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Make sure to wipe down pipettes between
style='color:windowtext;font-weight:normal;font-style:normal'>Make sure to wipe
+
solutions (with Ethanol)</span></span></p>
down pipettes between solutions (with Ethanol)<u><o:p></o:p></u></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Change tips every time you go between
style='color:windowtext;font-weight:normal;font-style:normal'>Change tips every
+
solutions</span></span></p>
time you go between solutions<u><o:p></o:p></u></span></span></p>
 
  
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span
-.25in;mso-list:l24 level1 lfo21'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span
+
</span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Solutions<o:p></o:p></span></u></span></p>
+
font-weight:normal;font-style:normal'>Solutions</span></u></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 lfo21'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Task: Make 0.3M KCl</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>Task: Make 0.3M
 
KCl<u><o:p></o:p></u></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span
-9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span
 
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>i.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span></span><![endif]><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>Salt (KCl): CAS 7447-40-7<u><o:p></o:p></u></span></span></p>
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>i.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>Salt (KCl): CAS 7447-40-7</span></span></p>
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span
-9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
</span>ii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>ii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span></span><![endif]><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>MW: 74.56g/mol<u><o:p></o:p></u></span></span></p>
+
font-style:normal'>MW: 74.56g/mol</span></span></p>
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span
-9.0pt;mso-list:l24 level3 lfo21'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
</span>iii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>iii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span></span><![endif]><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>2.2368 g / 100mL<u><o:p></o:p></u></span></span></p>
+
font-style:normal'>2.2368 g / 100mL</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><o:p>&nbsp;</o:p></span></span></p>
+
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
  
 
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
 
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
  style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt:
+
  style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+
border:none'>
  <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
+
  <tr>
 
   <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 
   <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
  background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto;
 
 
   padding:0in 5.4pt 0in 5.4pt'>
 
   padding:0in 5.4pt 0in 5.4pt'>
   <p class=MsoTitle style1 style='margin-right:26.1pt'>Plate Creation &amp; Cell
+
   <p class=MsoTitle style='margin-right:26.1pt'>Plate Creation &amp; Cell
   Culture</p> </td>
+
   Culture</p>
 +
  </td>
 
   <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 
   <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
  background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto;
 
 
   padding:0in 5.4pt 0in 5.4pt'>
 
   padding:0in 5.4pt 0in 5.4pt'>
   <p class=ExperimentNumber style1>Pg. 7</p> </td>
+
   <p class=ExperimentNumber>Pg. 7</p>
 +
  </td>
 
  </tr>
 
  </tr>
 
</table>
 
</table>
  
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span
-.25in;mso-list:l16 level1 lfo23'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span
+
</span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Steps for
+
font-weight:normal;font-style:normal'>Steps for creating a plate</span></u></span><span
creating a plate</span></u></span><span class=MsoIntenseEmphasis><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
style='color:windowtext;font-weight:normal;font-style:normal'>:<u><o:p></o:p></u></span></span></p>
+
font-style:normal'>:</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Dissolve 25g LB in 800mL of H<sub>2</sub>O</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>Dissolve 25g LB
 
in 800mL of H<sub>2</sub>O<u><o:p></o:p></u></span></span></p>
 
  
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>Bring the final
 
style='color:windowtext;font-weight:normal;font-style:normal'>Bring the final
volume to 1L in a 2L flask<o:p></o:p></span></span></p>
+
volume to 1L in a 2L flask</span></span></p>
  
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>Add 15g
 
style='color:windowtext;font-weight:normal;font-style:normal'>Add 15g
Bactoagar, cover with foil, autoclave for 20-30 minutes<o:p></o:p></span></span></p>
+
Bactoagar, cover with foil, autoclave for 20-30 minutes</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Bring back to lab, cool to 50</span></span><span
style='color:windowtext;font-weight:normal;font-style:normal'>Bring back to
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
lab, cool to 50</span></span><span class=MsoIntenseEmphasis><span
 
style='mso-bidi-font-family:Arial;color:windowtext;font-weight:normal;
 
 
font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span
 
font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>C </span></span><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>C </span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>while
 +
stirring</span></span></p>
 +
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'>while stirring</span></span><span
+
font-style:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-style:normal'><o:p></o:p></span></u></span></p>
+
font-weight:normal;font-style:normal'>Add100mg AMP &amp; let swirl until
 +
dissolved</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Pour it into the plates (about 2-3mm
style='color:windowtext;font-weight:normal;font-style:normal'>Add100mg AMP
+
deep)</span></span></p>
&amp; let swirl until dissolved<u><o:p></o:p></u></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>Pour it into the
 
plates (about 2-3mm deep)<u><o:p></o:p></u></span></span></p>
 
 
 
<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt:
 
-.25in;mso-list:l16 level1 lfo23'><![if !supportLists]><span
 
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><u><span
+
</span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Cell Culture</span></u></span><span
+
font-weight:normal;font-style:normal'>Cell Culture</span></u></span><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>:<u><o:p></o:p></u></span></span></p>
+
font-style:normal'>:</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Use solution of LB with AMP</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>Use solution of
 
LB with AMP<u><o:p></o:p></u></span></span></p>
 
  
 
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span
 
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>Ensure to keep the bags sealed to keep everything sterile <o:p></o:p></span></span></p>
+
font-style:normal'>Ensure to keep the bags sealed to keep everything sterile </span></span></p>
  
 
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span
 
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
font-style:normal'>*Note once a bag of plates has been opened, the whole thing
 
font-style:normal'>*Note once a bag of plates has been opened, the whole thing
must be used<o:p></o:p></span></span></p>
+
must be used</span></span></p>
  
 
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span
 
<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>The tubes allow the solution to be aerated<o:p></o:p></span></span></p>
+
font-style:normal'>The tubes allow the solution to be aerated</span></span></p>
  
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>The
+
style='color:windowtext;font-weight:normal'>The tubes with bacteria should look
tubes with bacteria should look cloudy upon visual inspection—can be confirmed
+
cloudy upon visual inspection—can be confirmed with cell density measurement
with cell density measurement (using the Spec)<o:p></o:p></span></span></p>
+
(using the Spec)</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Sterilize the autopipette, set to 1mL
style='color:windowtext;font-weight:normal;font-style:normal'>Sterilize the
+
(make a 2mL culture, however)</span></span></p>
autopipette, set to 1mL (make a 2mL culture, however)<o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>Add 2mL of the LB
 
style='color:windowtext;font-weight:normal;font-style:normal'>Add 2mL of the LB
(for a miniprep); you will have to add 1mL, two times<o:p></o:p></span></span></p>
+
(for a miniprep); you will have to add 1mL, two times</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Label one as the control and another as
style='color:windowtext;font-weight:normal;font-style:normal'>Label one as the
+
your experimental sample</span></span></p>
control and another as your experimental sample<o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>*Label the tubes
 
style='color:windowtext;font-weight:normal;font-style:normal'>*Label the tubes
with your Initials, Sample Name (control or experimental) and the date<o:p></o:p></span></span></p>
+
with your Initials, Sample Name (control or experimental) and the date</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Dip the culture scraper in ethanol and
style='color:windowtext;font-weight:normal;font-style:normal'>Dip the culture
+
flame lightly (2x) to sterilize</span></span></p>
scraper in ethanol and flame lightly (2x) to sterilize<o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>e.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>e.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Let the scraper cool for a few seconds</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>Let the scraper
 
cool for a few seconds<o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>f.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>f.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Scrape off a tiny bit of bacteria off of
style='color:windowtext;font-weight:normal;font-style:normal'>Scrape off a tiny
+
the plate and place the scraper into the tube and shake around to ensure the
bit of bacteria off of the plate and place the scraper into the tube and shake
+
bacteria breaks off into the tube</span></span></p>
around to ensure the bacteria breaks off into the tube<o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 lfo23'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>g.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>g.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>Set incubator at 37</span></span><span
style='color:windowtext;font-weight:normal;font-style:normal'>Set incubator at
 
37</span></span><span class=MsoIntenseEmphasis><span style='mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;font-style:normal'>°</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>C overnight and let shake gently for 12-14hrs<o:p></o:p></span></span></p>
+
font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>C overnight and
 +
let shake gently for 12-14hrs</span></span></p>
  
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
style='color:windowtext;font-weight:normal;font-style:normal'><o:p>&nbsp;</o:p></span></span></p>
+
style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
  
 
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
 
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
  style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt:
+
  style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+
border:none'>
  <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
+
  <tr>
 
   <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 
   <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
  background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto;
 
 
   padding:0in 5.4pt 0in 5.4pt'>
 
   padding:0in 5.4pt 0in 5.4pt'>
   <p class=MsoTitle style1 style='margin-right:26.1pt'>Transformation of E.coli with
+
   <p class=MsoTitle style='margin-right:26.1pt'>Transformation of E.coli with
   Recombinant DNA<o:p></o:p></p>  </td>
+
   Recombinant DNA</p>
 +
  </td>
 
   <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 
   <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
  background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto;
 
 
   padding:0in 5.4pt 0in 5.4pt'>
 
   padding:0in 5.4pt 0in 5.4pt'>
   <p class=ExperimentNumber style1>Pg. 9<o:p></o:p></p> </td>
+
   <p class=ExperimentNumber>Pg. 9</p>
 +
  </td>
 
  </tr>
 
  </tr>
 
</table>
 
</table>
Line 1,693: Line 473:
 
<p class=MsoNormal style='margin-left:.75in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:.75in'><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>*note, </span></span><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>*note, </span></span><span
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>the
mso-bidi-font-style:normal'>the heat shock</span></span><span
+
heat shock</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
+
font-weight:normal;font-style:normal'> to the tubes is very important for
font-style:normal'> to the tubes is very important for transcription to occur<o:p></o:p></span></span></p>
+
transcription to occur</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'>            </span><u>+
+
font-weight:normal;font-style:normal'>            <u>+ Controls</u>:</span></span></p>
Controls</u>:<o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_I13522
+
font-weight:normal;font-style:normal'>                        BBa_I13522 :
: pTetGFP<o:p></o:p></span></span></p>
+
pTetGFP</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_J45220<o:p></o:p></span></span></p>
+
font-weight:normal;font-style:normal'>                        BBa_J45220</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'>            </span><u>Group
+
font-weight:normal;font-style:normal'>            <u>Group 1:</u></span></span></p>
1:</u><o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_R0011<o:p></o:p></span></span></p>
+
font-weight:normal;font-style:normal'>                        BBa_R0011</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_J45219<o:p></o:p></span></span></p>
+
font-weight:normal;font-style:normal'>                        BBa_J45219</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'>            </span><u>Group
+
font-weight:normal;font-style:normal'>            <u>Group 2:</u></span></span></p>
2:</u><o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_I14032
+
font-weight:normal;font-style:normal'>                        BBa_I14032 /
/ BBa_R0051 (same)<o:p></o:p></span></span></p>
+
BBa_R0051 (same)</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_J06702<o:p></o:p></span></span></p>
+
font-weight:normal;font-style:normal'>                        BBa_J06702</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:1'>            </span><u>Group
+
font-weight:normal;font-style:normal'>            <u>Group 3:</u></span></span></p>
3:</u><o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_J04500<o:p></o:p></span></span></p>
+
font-weight:normal;font-style:normal'>                        BBa_J04500</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><span style='mso-tab-count:2'>                        </span>BBa_J04630
+
font-weight:normal;font-style:normal'>                        BBa_J04630 /
/ BBa_I13401<o:p></o:p></span></span></p>
+
BBa_I13401</span></span></p>
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><o:p>&nbsp;</o:p></span></span></p>
+
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
  
 
<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Rima
+
style='color:windowtext;font-weight:normal'>Rima &amp; I did the transformation
&amp; I did the transformation for Group 2:<o:p></o:p></span></span></p>
+
for Group 2:</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>BBa_I14032</span></span><span
+
font-weight:normal;font-style:normal'>BBa_I14032</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Name:
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name: </span></span><span
+
font-weight:normal;font-style:normal'>promoter P(Lac) IQ</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
font-style:normal'>promoter P(Lac) IQ</span></span><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Sequence:
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Sequence:
 
 
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
font-weight:normal;font-style:normal'>promoter</span></span><span
 
font-weight:normal;font-style:normal'>promoter</span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'> </span></span><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'> </span></span><span class=MsoIntenseEmphasis><span
+
font-style:normal'>(-35, -10 bases before coding)</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>(-35, -10 bases
 
before coding)</span></span><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Location</span></span><span
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>: 2007, Plate 2, Well 17K</span></span><span
+
font-style:normal'>: 2007, Plate 2, Well 17K</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>d.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Plasmid:
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Plasmid:
 
 
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'>pSB2K3</span></span><span
+
font-weight:normal;font-style:normal'>pSB2K3</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
 
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span
 
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>BBa_R0051</span></span><span
+
font-weight:normal;font-style:normal'>BBa_R0051</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Name:</span></span><span
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name:</span></span><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
font-style:normal'> promoter (lamda cl regulated)</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
+
font-style:normal'> promoter (lamda cl regulated)</span></span></p>
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Sequence:</span></span><span
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Sequence:</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'> R0051 promoter</span></span><span class=MsoIntenseEmphasis><span
+
font-style:normal'> R0051 promoter</span></span></p>
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Location:
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location:
 
 
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'>2007, Plate 1, Well 9C</span></span><span
+
font-weight:normal;font-style:normal'>2007, Plate 1, Well 9C</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
+
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><i>d.<span
 +
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></i><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Plasmid:</span></span><i>
 +
</i>pSB1A2</p>
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><i
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
style='mso-bidi-font-style:normal'><span style='mso-fareast-font-family:Arial;
 
mso-bidi-font-family:Arial'><span style='mso-list:Ignore'>d.<span
 
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span></i><![endif]><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'>Plasmid:</span></span><i style='mso-bidi-font-style:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
normal'> </i>pSB1A2<i style='mso-bidi-font-style:normal'><o:p></o:p></i></p>
 
 
 
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
 
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span
 
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>BBa_J06702</span></span><span
+
font-weight:normal;font-style:normal'>BBa_J06702</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Name:</span></span><span
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Name:</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'> mCherry, bacterial with RBS &amp; forward terminator</span></span><span
+
font-style:normal'> mCherry, bacterial with RBS &amp; forward terminator</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>Sequence</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'>: <o:p></o:p></span></span></p>
+
font-style:normal'>Sequence</span></span><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>: </span></span></p>
  
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
  style='margin-left:159.5pt;border-collapse:collapse;border:none;mso-border-alt:
+
  style='margin-left:159.5pt;border-collapse:collapse;border:none'>
solid windowtext .5pt;mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
+
  <tr>
  <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 
 
   <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt;
 
   <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt;
   mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
+
   padding:0in 5.4pt 0in 5.4pt'>
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;mso-bidi-font-style:normal'>RBS<o:p></o:p></span></span></p>
+
   font-weight:normal'>RBS</span></span></p>
 
   </td>
 
   </td>
 
   <td width=85 valign=top style='width:64.05pt;border:solid windowtext 1.0pt;
 
   <td width=85 valign=top style='width:64.05pt;border:solid windowtext 1.0pt;
   border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+
   border-left:none;padding:0in 5.4pt 0in 5.4pt'>
  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;mso-bidi-font-style:normal'>mCherry<o:p></o:p></span></span></p>
+
   font-weight:normal'>mCherry</span></span></p>
 
   </td>
 
   </td>
 
   <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt;
 
   <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt;
   border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+
   border-left:none;padding:0in 5.4pt 0in 5.4pt'>
  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;mso-bidi-font-style:normal'>Terminator<o:p></o:p></span></span></p>
+
   font-weight:normal'>Terminator</span></span></p>
 
   </td>
 
   </td>
 
   <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt;
 
   <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt;
   border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
+
   border-left:none;padding:0in 5.4pt 0in 5.4pt'>
  solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;mso-bidi-font-style:normal'>Terminator<o:p></o:p></span></span></p>
+
   font-weight:normal'>Terminator</span></span></p>
 
   </td>
 
   </td>
 
  </tr>
 
  </tr>
  <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
+
  <tr>
 
   <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt;
 
   <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt;
   border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
+
   border-top:none;padding:0in 5.4pt 0in 5.4pt'>
  padding:0in 5.4pt 0in 5.4pt'>
 
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;font-style:normal'>B0034<o:p></o:p></span></span></p>
+
   font-weight:normal;font-style:normal'>B0034</span></span></p>
 
   </td>
 
   </td>
 
   <td width=85 valign=top style='width:64.05pt;border-top:none;border-left:
 
   <td width=85 valign=top style='width:64.05pt;border-top:none;border-left:
 
   none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 
   none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
   mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+
   padding:0in 5.4pt 0in 5.4pt'>
  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;font-style:normal'>J06504<o:p></o:p></span></span></p>
+
   font-weight:normal;font-style:normal'>J06504</span></span></p>
 
   </td>
 
   </td>
 
   <td width=101 valign=top style='width:76.05pt;border-top:none;border-left:
 
   <td width=101 valign=top style='width:76.05pt;border-top:none;border-left:
 
   none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 
   none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
   mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+
   padding:0in 5.4pt 0in 5.4pt'>
  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;font-style:normal'>B0010<o:p></o:p></span></span></p>
+
   font-weight:normal;font-style:normal'>B0010</span></span></p>
 
   </td>
 
   </td>
 
   <td width=101 valign=top style='width:76.05pt;border-top:none;border-left:
 
   <td width=101 valign=top style='width:76.05pt;border-top:none;border-left:
 
   none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 
   none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
   mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
+
   padding:0in 5.4pt 0in 5.4pt'>
  mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
   <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
   font-weight:normal;font-style:normal'>B0012<o:p></o:p></span></span></p>
+
   font-weight:normal;font-style:normal'>B0012</span></span></p>
 
   </td>
 
   </td>
 
  </tr>
 
  </tr>
 
</table>
 
</table>
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Location:</span></span><span
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Location:</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'> 2007, Plate 2, Well 11D</span></span><span
+
font-style:normal'> 2007, Plate 2, Well 11D</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>d.<span
 +
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Plasmid:</span></span><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
+
font-style:normal'> pSB1A2</span></span></p>
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
 
style='mso-list:Ignore'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Plasmid:</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'> pSB1A2</span></span><span class=MsoIntenseEmphasis><span
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
 
 
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
 
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span
 
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Cells Used:
+
font-weight:normal;font-style:normal'>Cells Used: E.coli: JM109 (Promega);
E.coli: JM109 (Promega); Catalog #: L2001</span></span><span
+
Catalog #: L2001</span></span></p>
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;mso-bidi-font-style:normal'><o:p>&nbsp;</o:p></span></span></p>
+
font-weight:normal'>&nbsp;</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l11 level1 lfo30'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>3 Plates Made:</span></span><span
+
font-weight:normal;font-style:normal'>3 Plates Made:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span
 +
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
+
font-style:normal'>Control: 100&#956;L LB / Cell</span></span></p>
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-style:normal'>Test Plate 1: 100&#956;L LB/ Cell</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>Control: 100&#956;L
 
LB / Cell</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
+
style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
style='mso-list:Ignore'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-style:normal'>Test Plate 2: 1&#956;L cell : 99&#956; H<sub>2</sub>O</span></span></p>
style='color:windowtext;font-weight:normal;font-style:normal'>Test Plate 1: 100&#956;L
 
LB/ Cell</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
 
 
<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in;mso-list:l11 level2 lfo30'><![if !supportLists]><span
 
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
 
Arial;color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><span
 
style='mso-list:Ignore'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'>Test Plate 2: 1&#956;L
 
cell : 99&#956; H<sub>2</sub>O</span></span><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'><o:p></o:p></span></span></p>
 
  
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
font-weight:normal;font-style:normal'><o:p>&nbsp;</o:p></span></span></p>
+
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
  
 
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Actual
+
style='color:windowtext;font-weight:normal'>Actual Transformation Procedure:</span></span></p>
Transformation Procedure:</span></span><span class=MsoIntenseEmphasis><span
 
style='color:windowtext;font-weight:normal;font-style:normal'><o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level1 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>If using DNA from
+
font-weight:normal;font-style:normal'>If using DNA from the registry, dilute
the registry, dilute with 9&#956;L of H<sub>2</sub>O. If using our own DNA, use
+
with 9&#956;L of H<sub>2</sub>O. If using our own DNA, use 200ng. </span></span></p>
200ng. <o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt;mso-list:l26 level3 lfo31'><![if !supportLists]><span
+
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt'><span
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
font-weight:normal;font-style:normal'>inject the water directly into the well,
style='color:windowtext;font-weight:normal;font-style:normal'>inject the water
+
titerate, and then pipette it out</span></span></p>
directly into the well, titerate, and then pipette it out<o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Thaw 100&#956;L
+
font-weight:normal;font-style:normal'>Thaw 100&#956;L competent cells on ice.
competent cells on ice. (50</span></span> <span class=MsoIntenseEmphasis><span
+
(50</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>&#956;L with our
+
font-weight:normal;font-style:normal'>&#956;L with our own/ 100</span></span> <span
own/ 100</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-weight:normal;font-style:normal'>&#956; if commercial cells used)<o:p></o:p></span></span></p>
+
font-style:normal'>&#956; if commercial cells used)</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Mix competent
+
font-weight:normal;font-style:normal'>Mix competent cells with 50</span></span>
cells with 50</span></span> <span class=MsoIntenseEmphasis><span
+
<span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
style='color:windowtext;font-weight:normal;font-style:normal'>&#956; CaCl<sub>2</sub>
+
font-style:normal'>&#956; CaCl<sub>2</sub> (only if using our own cells)</span></span></p>
(only if using our own cells)<o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Add 4</span></span>
+
font-weight:normal;font-style:normal'>Add 4</span></span> <span
<span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
font-style:normal'>&#956;L of registry DNA, or 200ng of our own DNA to the
 
font-style:normal'>&#956;L of registry DNA, or 200ng of our own DNA to the
cells. <o:p></o:p></span></span></p>
+
cells. </span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Put in the ice
+
font-weight:normal;font-style:normal'>Put in the ice bath for about 20 minutes.
bath for about 20 minutes. <o:p></o:p></span></span></p>
+
</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>VI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>VI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Remove and
+
font-weight:normal;font-style:normal'>Remove and immediately put in the hot
immediately put in the hot water bath (42</span></span><span
+
water bath (42</span></span><span class=MsoIntenseEmphasis><span
class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext;
+
style='color:windowtext;font-weight:normal;font-style:normal'>°</span></span><span
font-weight:normal;font-style:normal'>°</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>C) to heat shock for 45-60 sec.<o:p></o:p></span></span></p>
+
font-style:normal'>C) to heat shock for 45-60 sec.</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>VII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>VII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Place back in the
+
font-weight:normal;font-style:normal'>Place back in the ice bath for 2 minutes.
ice bath for 2 minutes. <o:p></o:p></span></span></p>
+
</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>VIII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>VIII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Add 900 &#956;L
+
font-weight:normal;font-style:normal'>Add 900 &#956;L ice cold, antibiotic free
ice cold, antibiotic free LB to the cells<o:p></o:p></span></span></p>
+
LB to the cells</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>IX.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>IX.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Incubate at 37</span></span><span
+
font-weight:normal;font-style:normal'>Incubate at 37</span></span><span
class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext;
 
font-weight:normal;font-style:normal'>°</span></span><span
 
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>C for 90 minutes in the shaking incubator.<o:p></o:p></span></span></p>
+
font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>C for 90 minutes
 +
in the shaking incubator.</span></span></p>
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>X.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>X.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Dilute as desired,
+
font-weight:normal;font-style:normal'>Dilute as desired, make the necessary
make the necessary controls, and plate cells using 100 &#956;L / plate. (200 if
+
controls, and plate cells using 100 &#956;L / plate. (200 if not commercial)</span></span></p>
not commercial)<o:p></o:p></span></span></p>
 
  
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in;mso-text-indent-alt:
+
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
-.25in;mso-list:l26 level2 lfo31'><![if !supportLists]><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
 
style='mso-list:Ignore'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
 
</span>XI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 
</span>XI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
style='color:windowtext;font-weight:normal;font-style:normal'>Incubate upside
+
font-weight:normal;font-style:normal'>Incubate upside down (so that the label
down (so that the label you wrote on it is face up) at 37</span></span><span
+
you wrote on it is face up) at 37</span></span><span class=MsoIntenseEmphasis><span
class=MsoIntenseEmphasis><span style='mso-bidi-font-family:Arial;color:windowtext;
+
style='color:windowtext;font-weight:normal;font-style:normal'>°</span></span><span
font-weight:normal;font-style:normal'>°</span></span><span
+
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>C for 18-24 hours, then refrigerate. </span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt'><span
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
font-style:normal'>C for 18-24 hours, then refrigerate. <o:p></o:p></span></span></p>
+
font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>The controls should not have bacteria
 +
growing on them!</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
  
<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt;mso-list:l26 level3 lfo31'><![if !supportLists]><span
+
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
class=MsoIntenseEmphasis><span style='mso-fareast-font-family:Arial;mso-bidi-font-family:
+
style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
Arial;color:windowtext;font-weight:normal;font-style:normal'><span
+
border:none'>
style='mso-list:Ignore'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
<tr>
</span></span></span></span><![endif]><span class=MsoIntenseEmphasis><span
+
  <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
style='color:windowtext;font-weight:normal;font-style:normal'>The controls
+
  padding:0in 5.4pt 0in 5.4pt'>
should not have bacteria growing on them!<o:p></o:p></span></span></p>
+
  <p class=MsoTitle style='margin-right:26.1pt'>Miniprep</p>
 +
  </td>
 +
  <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 +
  padding:0in 5.4pt 0in 5.4pt'>
 +
  <p class=ExperimentNumber>Pg. 12</p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>*Start with
 +
Parts: E0241 (x3), R0010, R0040, C0040 from the registry*</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>E0241: PoPs to GFP Converter</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext;
 +
font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>RBS: B0032</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext;
 +
font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>GFP: E0040</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext;
 +
font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Terminator: B0016</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>R0010: Promoter Lac I (Regulator) </span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>R0040: Promoter (tetR, negative)</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>C0040: Tetracycline repressor from
 +
transposon Tn10 (+LVA) [note that +LVA is a degradation tag]</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>Look out for:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>            </span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>Control should not be cloudy at all, culture should have gas
 +
bubbles, should be cloudy, especially at the bottom</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            *If
 +
you start seeing sediment at the bottom, the cells </span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>may</span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'> have lysed and let proteins into the tube*</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>            </span></span></p>
 +
 
 +
<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>Procedure (General):</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>Use the QIAGEN
 +
Kit to separate the plasmid DNA (so it should not have much chromosomal DNA)</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Grow cells for 12 to 14 hours</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Suspend cells in </span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>i.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>buffer 1 (with glucose &amp; EDTA)</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>ii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>buffer 2 (SDS-detergent, NaOH) [SDS gets rid of the membrane
 +
and NaOH denatures the plasmid and DNA]</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>iii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>buffer 3 (K Acetate / Acetic Acid)</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>-things become an insoluble precipitate</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>-renature the DNA/plasmid</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>-only the plasmid DNA will renature properly</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>-the plasmid DNA remains soluble &amp; the chromosomal DNA
 +
will become the precipitate</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Columns: use EtOH to extract DNA</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>Procedure (Specific):</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            *make
 +
sure to check your buffers for salt precipitation—this may indicate a change in
 +
concentration*</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Get solution and put 1.5mL into an Eppendorf
 +
tube to centrifuge </span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>-after
 +
centrifuging for 30 seconds @ 13k RPM, remove the supernatant</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>-add more
 +
solution, and centrifuge again using same procedure</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Add 250</span></span> <span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>&#956;L PI to tube after the supernatant is </span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>fully
 +
removed</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Add 250 &#956;L P2, and invert 4 to 6
 +
times</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>-immediately add
 +
350</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>&#956;L N3, and invert again 4 to 6 times</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Dispose of the flow through, add 0.5mL of
 +
buffer PB (that is </span></span><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>milliliter not microliter!)</span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'> then centrifuge for 30-60 seconds</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Add 750</span></span> <span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>&#956;L PE, then centrifuge again, remove the waste,
 +
centrifuge again</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>VI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Add 50</span></span> <span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>&#956;L BE buffer</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>VII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Measure the DNA concentration using
 +
Nanodrop</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>VIII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Ethanol Precipitation </span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>(see
 +
separate procedure)</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal'>&nbsp;</span></span></p>
 +
 
 +
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
 +
style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
 +
border:none'>
 +
<tr>
 +
  <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 +
  padding:0in 5.4pt 0in 5.4pt'>
 +
  <p class=MsoTitle style='margin-right:26.1pt'>Ethanol Precipitation (for
 +
  &gt;200bp)</p>
 +
  </td>
 +
  <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 +
  padding:0in 5.4pt 0in 5.4pt'>
 +
  <p class=ExperimentNumber>Pg. 15</p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>            *used to purify the DNA
 +
(really small fragments) or to increase the concentration of it</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Add:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-1.5in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>i.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>3 volumes of COLD 100% EtOH</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-1.5in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>ii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>1/10 volume of 3M sodium acetate</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-1.5in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>iii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>1 &#956;L GlycoBlue</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Incubate for 1 hr @ 80</span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>C</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Centrifuge for 30min @ 0</span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>C</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Remove supernatant</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Air Dry</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span>VI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Resuspend in H<sub>2</sub>O</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>Specific Procedure Example:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            For
 +
50 &#956;L:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>                        Add
 +
    150 &#956;L EtOH</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>                                    20
 +
&#956;L NaAc.</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            For
 +
100 &#956;L:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>                        Add    300
 +
&#956;L EtOH</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>                                    40
 +
&#956;L NaAc.</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
 +
style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
 +
border:none'>
 +
<tr>
 +
  <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 +
  padding:0in 5.4pt 0in 5.4pt'>
 +
  <p class=MsoTitle style='margin-right:26.1pt'>Gel Electrophoresis</p>
 +
  </td>
 +
  <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 +
  padding:0in 5.4pt 0in 5.4pt'>
 +
  <p class=ExperimentNumber>Pg. 16</p>
 +
  </td>
 +
</tr>
 +
</table>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>            *applying an electric field
 +
over the gel will separate fragments by size</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>            *compare the movement of the
 +
DNA to the ladder</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>            </span></span><span
 +
class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal'>*ensure
 +
that the well is on the negative side!*</span></u></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>DNA has a (-) charge because of the
 +
phosphate groups</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>moves toward the anode (+) and away from
 +
the cathode (-)</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>DNA is separated by size, distance ~ 1 /
 +
molecular weight</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext;
 +
font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>100bp-50kbp</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>Gel acts as a sieve: 0.3% large fragments
 +
to 2% small fragments</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span
 +
class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext;
 +
font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;
 +
</span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>after the gel is made, put it in the
 +
microwave for 1minute, then </span></span><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>cool the bottle in H­<sub>2</sub>O,
 +
and take the lid off</span></span></p>
 +
 
 +
<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 +
font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>Casting Gel:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            1%
 +
Agarose, 0.5g Agarose</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            50mL
 +
TBE + SYBR Safe</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>Dilute the 5xTBE
 +
to 0.5x</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>Add 2 &#956;L of
 +
the loading dye!</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>Ladder Formula:</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal'>            </span></span><span
 +
class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;
 +
font-style:normal'>1 &#956;L 1kbase ladder</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            2 &#956;L
 +
BlueJuice Dye</span></span></p>
 +
 
 +
<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span
 +
style='color:windowtext;font-weight:normal;font-style:normal'>            7 &#956;L
 +
H<sub>2</sub>O</span></span></p>
  
 
</div>
 
</div>

Revision as of 22:13, 25 February 2008

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</head>

<body lang=EN-US link=blue vlink=purple>

<div class=Section1>

<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0

style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
border:none'>
<tr>
 <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoTitle style='margin-right:26.1pt'>Pipetting &amp; Making
 Solutions</p>
 </td>
 <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=ExperimentNumber>Pg. 6</p>
 </td>
</tr>

</table>

<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext; font-weight:normal;font-style:normal'>Pipetting</span></u></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Learned to use pipettes properly</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Make sure to wipe down pipettes between solutions (with Ethanol)</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Change tips every time you go between solutions</span></span></p>

<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext; font-weight:normal;font-style:normal'>Solutions</span></u></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Task: Make 0.3M KCl</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>i.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Salt (KCl): CAS 7447-40-7</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>ii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>MW: 74.56g/mol</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>iii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>2.2368 g / 100mL</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0

style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
border:none'>
<tr>
 <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoTitle style='margin-right:26.1pt'>Plate Creation &amp; Cell
 Culture</p>
 </td>
 <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=ExperimentNumber>Pg. 7</p>
 </td>
</tr>

</table>

<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext; font-weight:normal;font-style:normal'>Steps for creating a plate</span></u></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>:</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Dissolve 25g LB in 800mL of H<sub>2</sub>O</span></span></p>

<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Bring the final volume to 1L in a 2L flask</span></span></p>

<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add 15g Bactoagar, cover with foil, autoclave for 20-30 minutes</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Bring back to lab, cool to 50</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>C </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>while stirring</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add100mg AMP &amp; let swirl until dissolved</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Pour it into the plates (about 2-3mm deep)</span></span></p>

<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext; font-weight:normal;font-style:normal'>Cell Culture</span></u></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>:</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Use solution of LB with AMP</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Ensure to keep the bags sealed to keep everything sterile </span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>*Note once a bag of plates has been opened, the whole thing must be used</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>The tubes allow the solution to be aerated</span></span></p>

<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>The tubes with bacteria should look cloudy upon visual inspection—can be confirmed with cell density measurement (using the Spec)</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Sterilize the autopipette, set to 1mL (make a 2mL culture, however)</span></span></p>

<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add 2mL of the LB (for a miniprep); you will have to add 1mL, two times</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Label one as the control and another as your experimental sample</span></span></p>

<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>*Label the tubes with your Initials, Sample Name (control or experimental) and the date</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Dip the culture scraper in ethanol and flame lightly (2x) to sterilize</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>e.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Let the scraper cool for a few seconds</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>f.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Scrape off a tiny bit of bacteria off of the plate and place the scraper into the tube and shake around to ensure the bacteria breaks off into the tube</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>g.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Set incubator at 37</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>C overnight and let shake gently for 12-14hrs</span></span></p>

<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0

style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
border:none'>
<tr>
 <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoTitle style='margin-right:26.1pt'>Transformation of E.coli with
 Recombinant DNA</p>
 </td>
 <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=ExperimentNumber>Pg. 9</p>
 </td>
</tr>

</table>

<p class=MsoNormal style='margin-left:.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>*note, </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>the heat shock</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> to the tubes is very important for transcription to occur</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> <u>+ Controls</u>:</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_I13522 : pTetGFP</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_J45220</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> <u>Group 1:</u></span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_R0011</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_J45219</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> <u>Group 2:</u></span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_I14032 / BBa_R0051 (same)</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_J06702</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> <u>Group 3:</u></span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_J04500</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> BBa_J04630 / BBa_I13401</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Rima &amp; I did the transformation for Group 2:</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>BBa_I14032</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Name: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>promoter P(Lac) IQ</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Sequence: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>promoter</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'> </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>(-35, -10 bases before coding)</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Location</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>: 2007, Plate 2, Well 17K</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Plasmid: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>pSB2K3</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>BBa_R0051</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Name:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> promoter (lamda cl regulated)</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Sequence:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> R0051 promoter</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Location: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>2007, Plate 1, Well 9C</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><i>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></i><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Plasmid:</span></span><i> </i>pSB1A2</p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>BBa_J06702</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Name:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> mCherry, bacterial with RBS &amp; forward terminator</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Sequence</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>: </span></span></p>

<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0

style='margin-left:159.5pt;border-collapse:collapse;border:none'>
<tr>
 <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal'>RBS</span></span></p>
 </td>
 <td width=85 valign=top style='width:64.05pt;border:solid windowtext 1.0pt;
 border-left:none;padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal'>mCherry</span></span></p>
 </td>
 <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt;
 border-left:none;padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal'>Terminator</span></span></p>
 </td>
 <td width=101 valign=top style='width:76.05pt;border:solid windowtext 1.0pt;
 border-left:none;padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal'>Terminator</span></span></p>
 </td>
</tr>
<tr>
 <td width=70 valign=top style='width:52.8pt;border:solid windowtext 1.0pt;
 border-top:none;padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>B0034</span></span></p>
 </td>
 <td width=85 valign=top style='width:64.05pt;border-top:none;border-left:
 none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>J06504</span></span></p>
 </td>
 <td width=101 valign=top style='width:76.05pt;border-top:none;border-left:
 none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>B0010</span></span></p>
 </td>
 <td width=101 valign=top style='width:76.05pt;border-top:none;border-left:
 none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>B0012</span></span></p>
 </td>
</tr>

</table>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Location:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> 2007, Plate 2, Well 11D</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>d.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Plasmid:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> pSB1A2</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Cells Used: E.coli: JM109 (Promega); Catalog #: L2001</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>3 Plates Made:</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Control: 100&#956;L LB / Cell</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>b.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Test Plate 1: 100&#956;L LB/ Cell</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>c.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Test Plate 2: 1&#956;L cell : 99&#956; H<sub>2</sub>O</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Actual Transformation Procedure:</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>If using DNA from the registry, dilute with 9&#956;L of H<sub>2</sub>O. If using our own DNA, use 200ng. </span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>inject the water directly into the well, titerate, and then pipette it out</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Thaw 100&#956;L competent cells on ice. (50</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&#956;L with our own/ 100</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>&#956; if commercial cells used)</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Mix competent cells with 50</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>&#956; CaCl<sub>2</sub> (only if using our own cells)</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add 4</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>&#956;L of registry DNA, or 200ng of our own DNA to the cells. </span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Put in the ice bath for about 20 minutes. </span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>VI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Remove and immediately put in the hot water bath (42</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>C) to heat shock for 45-60 sec.</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>VII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Place back in the ice bath for 2 minutes. </span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>VIII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add 900 &#956;L ice cold, antibiotic free LB to the cells</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>IX.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Incubate at 37</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>C for 90 minutes in the shaking incubator.</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>X.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Dilute as desired, make the necessary controls, and plate cells using 100 &#956;L / plate. (200 if not commercial)</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>XI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Incubate upside down (so that the label you wrote on it is face up) at 37</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>C for 18-24 hours, then refrigerate. </span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-9.0pt'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>a.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>The controls should not have bacteria growing on them!</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0

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<tr>
 <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoTitle style='margin-right:26.1pt'>Miniprep</p>
 </td>
 <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
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 <p class=ExperimentNumber>Pg. 12</p>
 </td>
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</table>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>*Start with Parts: E0241 (x3), R0010, R0040, C0040 from the registry*</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>E0241: PoPs to GFP Converter</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext; font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>RBS: B0032</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext; font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>GFP: E0040</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext; font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Terminator: B0016</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>R0010: Promoter Lac I (Regulator) </span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>R0040: Promoter (tetR, negative)</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>C0040: Tetracycline repressor from transposon Tn10 (+LVA) [note that +LVA is a degradation tag]</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Look out for:</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'> </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>Control should not be cloudy at all, culture should have gas bubbles, should be cloudy, especially at the bottom</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> *If you start seeing sediment at the bottom, the cells </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>may</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> have lysed and let proteins into the tube*</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> </span></span></p>

<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Procedure (General):</span></span></p>

<p class=MsoNormal style='margin-left:1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Use the QIAGEN Kit to separate the plasmid DNA (so it should not have much chromosomal DNA)</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Grow cells for 12 to 14 hours</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Suspend cells in </span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>i.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>buffer 1 (with glucose &amp; EDTA)</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>ii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>buffer 2 (SDS-detergent, NaOH) [SDS gets rid of the membrane and NaOH denatures the plasmid and DNA]</span></span></p>

<p class=MsoNormal style='margin-left:1.75in;text-indent:-1.75in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>iii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>buffer 3 (K Acetate / Acetic Acid)</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>-things become an insoluble precipitate</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>-renature the DNA/plasmid</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>-only the plasmid DNA will renature properly</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>-the plasmid DNA remains soluble &amp; the chromosomal DNA will become the precipitate</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Columns: use EtOH to extract DNA</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Procedure (Specific):</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> *make sure to check your buffers for salt precipitation—this may indicate a change in concentration*</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Get solution and put 1.5mL into an Eppendorf tube to centrifuge </span></span></p>

<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>-after centrifuging for 30 seconds @ 13k RPM, remove the supernatant</span></span></p>

<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>-add more solution, and centrifuge again using same procedure</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add 250</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>&#956;L PI to tube after the supernatant is </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>fully removed</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add 250 &#956;L P2, and invert 4 to 6 times</span></span></p>

<p class=MsoNormal style='margin-left:1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>-immediately add 350</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&#956;L N3, and invert again 4 to 6 times</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Dispose of the flow through, add 0.5mL of buffer PB (that is </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>milliliter not microliter!)</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> then centrifuge for 30-60 seconds</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add 750</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>&#956;L PE, then centrifuge again, remove the waste, centrifuge again</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>VI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add 50</span></span> <span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>&#956;L BE buffer</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>VII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Measure the DNA concentration using Nanodrop</span></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-1.25in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>VIII.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Ethanol Precipitation </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>(see separate procedure)</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal'>&nbsp;</span></span></p>

<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0

style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
border:none'>
<tr>
 <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoTitle style='margin-right:26.1pt'>Ethanol Precipitation (for
 &gt;200bp)</p>
 </td>
 <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=ExperimentNumber>Pg. 15</p>
 </td>
</tr>

</table>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> *used to purify the DNA (really small fragments) or to increase the concentration of it</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>I.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Add:</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>i.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>3 volumes of COLD 100% EtOH</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>ii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>1/10 volume of 3M sodium acetate</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-1.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>iii.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>1 &#956;L GlycoBlue</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>II.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Incubate for 1 hr @ 80</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>C</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>III.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Centrifuge for 30min @ 0</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>°</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>C</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>IV.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Remove supernatant</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>V.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Air Dry</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-1.0in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'><span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span>VI.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Resuspend in H<sub>2</sub>O</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Specific Procedure Example:</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> For 50 &#956;L:</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> Add

   150 &#956;L EtOH</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> 20 &#956;L NaAc.</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> For 100 &#956;L:</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> Add 300 &#956;L EtOH</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> 40 &#956;L NaAc.</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0

style='margin-left:5.4pt;background:#CCCCCC;border-collapse:collapse;
border:none'>
<tr>
 <td width=488 style='width:365.7pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoTitle style='margin-right:26.1pt'>Gel Electrophoresis</p>
 </td>
 <td width=136 style='width:102.3pt;border:none;border-bottom:solid windowtext 2.25pt;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=ExperimentNumber>Pg. 16</p>
 </td>
</tr>

</table>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> *applying an electric field over the gel will separate fragments by size</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> *compare the movement of the DNA to the ladder</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'> </span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal'>*ensure that the well is on the negative side!*</span></u></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>DNA has a (-) charge because of the phosphate groups</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>moves toward the anode (+) and away from the cathode (-)</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>DNA is separated by size, distance ~ 1 / molecular weight</span></span></p>

<p class=MsoNormal style='margin-left:1.5in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Wingdings;color:windowtext; font-weight:normal;font-style:normal'>§<span style='font:7.0pt "Times New Roman"'>&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>100bp-50kbp</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>Gel acts as a sieve: 0.3% large fragments to 2% small fragments</span></span></p>

<p class=MsoNormal style='margin-left:1.0in;text-indent:-.25in'><span class=MsoIntenseEmphasis><span style='font-family:Symbol;color:windowtext; font-weight:normal;font-style:normal'>®<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp; </span></span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>after the gel is made, put it in the microwave for 1minute, then </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>cool the bottle in H­<sub>2</sub>O, and take the lid off</span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Casting Gel:</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> 1% Agarose, 0.5g Agarose</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> 50mL TBE + SYBR Safe</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Dilute the 5xTBE to 0.5x</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Add 2 &#956;L of the loading dye!</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>&nbsp;</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'>Ladder Formula:</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal'> </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>1 &#956;L 1kbase ladder</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> 2 &#956;L BlueJuice Dye</span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'> 7 &#956;L H<sub>2</sub>O</span></span></p>

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