User:Neil J. Parikh/Lab Notebook

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</head>

<body lang=EN-US link=blue vlink=purple style='tab-interval:.5in'>

<div class=Section1>

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

style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt:
solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
 <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'>
 <p class=MsoTitle style1 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;
 background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=ExperimentNumber style1>Pg. 6</p>  </td>
</tr>

</table>

<p class=MsoNormal style='margin-left:.75in;text-indent:-.75in;mso-text-indent-alt: -.25in;mso-list:l24 level1 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; </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 style='color:windowtext;font-weight:normal;font-style:normal'>Pipetting<o:p></o:p></span></u></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 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'>a.<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'>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 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'>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'>Make sure to wipe 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 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'>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'>Change tips every 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: -.25in;mso-list:l24 level1 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; </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 style='color:windowtext;font-weight:normal;font-style:normal'>Solutions<o:p></o:p></span></u></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l24 level2 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'>a.<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'>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: -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; font-style:normal'>Salt (KCl): CAS 7447-40-7<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: -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; </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; font-style:normal'>MW: 74.56g/mol<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: -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; </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; font-style:normal'>2.2368 g / 100mL<u><o:p></o:p></u></span></span></p>

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

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

style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt:
solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
 <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'>
 <p class=MsoTitle style1 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;
 background:#333333;mso-shading:windowtext;mso-pattern:gray-80 auto;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=ExperimentNumber style1>Pg. 7</p>  </td>
</tr>

</table>

<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;&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 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'>:<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 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'>a.<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'>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 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>

<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<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 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'>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'>Bring back to 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 style='color:windowtext;font-weight:normal;font-style:normal'>C </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; mso-bidi-font-style:normal'>while stirring</span></span><span class=MsoIntenseEmphasis><u><span style='color:windowtext;font-weight:normal; font-style:normal'><o:p></o:p></span></u></span></p>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 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'>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'>Add100mg AMP &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 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'>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></span></span></span><![endif]><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'>:<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 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'>a.<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'>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 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>

<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<o:p></o:p></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<o:p></o:p></span></span></p>

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

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 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'>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'>Label one as the 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 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>

<p class=MsoNormal style='margin-left:1.25in;text-indent:-.25in;mso-list:l16 level2 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'>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'>Dip the culture 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 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'>e.<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'>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 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'>f.<span style='font:7.0pt "Times New Roman"'>&nbsp;&nbsp;&nbsp;&nbsp; </span></span></span></span><![endif]><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<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 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'>g.<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'>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; font-style:normal'>C overnight and let shake gently for 12-14hrs<o:p></o:p></span></span></p>

<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>

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

style='margin-left:5.4pt;border-collapse:collapse;border:none;mso-border-bottom-alt:
solid windowtext .5pt;mso-yfti-tbllook:191;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
 <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'>
 <p class=MsoTitle style1 style='margin-right:26.1pt'>Transformation of E.coli with
 Recombinant DNA<o:p></o:p></p>  </td>
 <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'>
 <p class=ExperimentNumber style1>Pg. 9<o:p></o:p></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; mso-bidi-font-style: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<o:p></o:p></span></span></p>

<p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'><span style='mso-tab-count:1'> </span><u>+ Controls</u>:<o:p></o:p></span></span></p>

<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

pTetGFP<o:p></o:p></span></span></p>

<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>

<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 1:</u><o:p></o:p></span></span></p>

<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>

<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>

<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 2:</u><o:p></o:p></span></span></p>

<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 / BBa_R0051 (same)<o:p></o:p></span></span></p>

<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>

<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 3:</u><o:p></o:p></span></span></p>

<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>

<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 / BBa_I13401<o:p></o:p></span></span></p>

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

<p class=MsoNormal style='text-indent:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Rima &amp; I did the transformation for Group 2:<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;&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 style='color:windowtext;font-weight:normal;font-style:normal'>BBa_I14032</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'>a.<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'>Name: </span></span><span 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 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'>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;mso-bidi-font-style: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; mso-bidi-font-style:normal'> </span></span><span class=MsoIntenseEmphasis><span 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 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;mso-bidi-font-style:normal'>Location</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'>: 2007, Plate 2, Well 17K</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'>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;font-style:normal'>pSB2K3</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:1.25in;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></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>BBa_R0051</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'>a.<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'>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; 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'>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;mso-bidi-font-style:normal'>Sequence:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> R0051 promoter</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;mso-bidi-font-style:normal'>Location: </span></span><span class=MsoIntenseEmphasis><span style='color:windowtext; font-weight:normal;font-style:normal'>2007, Plate 1, Well 9C</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]><i 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; mso-bidi-font-style:normal'>Plasmid:</span></span><i style='mso-bidi-font-style: 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></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>BBa_J06702</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'>a.<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'>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><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'>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; mso-bidi-font-style:normal'>: <o:p></o:p></span></span></p>

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

style='margin-left:159.5pt;border-collapse:collapse;border:none;mso-border-alt:
solid windowtext .5pt;mso-yfti-tbllook:1184;mso-padding-alt:0in 5.4pt 0in 5.4pt'>
<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
 <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'>
 <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>
 </td>
 <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:
 solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 <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>
 </td>
 <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:
 solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 <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>
 </td>
 <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:
 solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 <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>
 </td>
</tr>
<tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
 <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;
 padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>B0034<o:p></o:p></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;
 mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>J06504<o:p></o:p></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;
 mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>B0010<o:p></o:p></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;
 mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
 mso-border-alt:solid windowtext .5pt;padding:0in 5.4pt 0in 5.4pt'>
 <p class=MsoNormal><span class=MsoIntenseEmphasis><span style='color:windowtext;
 font-weight:normal;font-style:normal'>B0012<o:p></o:p></span></span></p>
 </td>
</tr>

</table>

<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;mso-bidi-font-style:normal'>Location:</span></span><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal; font-style:normal'> 2007, Plate 2, Well 11D</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'>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; font-style:normal'> pSB1A2</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:1.25in;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></span></span></span><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>Cells Used: E.coli: JM109 (Promega); Catalog #: L2001</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; font-weight:normal;mso-bidi-font-style:normal'><o:p>&nbsp;</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;&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 style='color:windowtext;font-weight:normal;font-style:normal'>3 Plates Made:</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'>a.<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'>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 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'>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'>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; font-weight:normal;font-style:normal'><o:p>&nbsp;</o:p></span></span></p>

<p class=MsoNormal style='margin-left:.5in'><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;mso-bidi-font-style:normal'>Actual 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: -.25in;mso-list:l26 level1 lfo31'><![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; </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 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. <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 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'>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><![endif]><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<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:l26 level2 lfo31'><![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></span></span></span><![endif]><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)<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:l26 level2 lfo31'><![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></span></span></span><![endif]><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)<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:l26 level2 lfo31'><![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></span></span></span><![endif]><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. <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:l26 level2 lfo31'><![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>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 style='color:windowtext;font-weight:normal;font-style:normal'>Put in the ice bath for about 20 minutes. <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:l26 level2 lfo31'><![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>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 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='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; font-style:normal'>C) to heat shock for 45-60 sec.<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:l26 level2 lfo31'><![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; </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 style='color:windowtext;font-weight:normal;font-style:normal'>Place back in the ice bath for 2 minutes. <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:l26 level2 lfo31'><![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; </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 style='color:windowtext;font-weight:normal;font-style:normal'>Add 900 &#956;L ice cold, antibiotic free LB to the cells<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:l26 level2 lfo31'><![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>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 style='color:windowtext;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; font-style:normal'>C for 90 minutes in the shaking incubator.<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:l26 level2 lfo31'><![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>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 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)<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:l26 level2 lfo31'><![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>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 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='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; font-style:normal'>C for 18-24 hours, then refrigerate. <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 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'>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><![endif]><span class=MsoIntenseEmphasis><span style='color:windowtext;font-weight:normal;font-style:normal'>The controls should not have bacteria growing on them!<o:p></o:p></span></span></p>

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