Difference between revisions of "Haynes Lab:Notebook/Engineering PC-TFs/2014/08/01"

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|style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span>
 
|style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span>
|style="background-color: #800000" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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|style="background-color: #800000" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
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| colspan="2"|
 
| colspan="2"|
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| bgcolor=#cfcfcf | Volume
 
| bgcolor=#cfcfcf | Volume
 
| rowspan="9" |  
 
| rowspan="9" |  
<u>Expected:</u>
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<u><b>Expected:</b></u>
 
<br>
 
<br>
BL09 Ligation Colonies 1-3[XbaI/NruI] (Well: 2-4)<br>  
+
<u>BL09 Ligation Colonies 1-3[XbaI/NruI] (Well: 2-4)</u><br>  
 
Forward insertion: 710, 5921 <br>
 
Forward insertion: 710, 5921 <br>
 
Reverse insertion: 2141, 4490 <br>
 
Reverse insertion: 2141, 4490 <br>
 
No insertion: 710, 4490<br> <br>
 
No insertion: 710, 4490<br> <br>
CMV/MV9 with [XbaI/NruI] (Well: 5) <br>
+
<u>CMV/MV9 with [XbaI/NruI] (Well: 5)</u> <br>
 
710, 4490<br><br>
 
710, 4490<br><br>
CMV/MV9 [XbaI](Well: 6) <br>
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<u>CMV/MV9 [XbaI](Well: 6)</u> <br>
 
5200<br><br>
 
5200<br><br>
Dephos CMV/MV9 (Well: 7) <br>
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<u>Dephos CMV/MV9 (Well: 7)</u> <br>
 
5200
 
5200
 
| rowspan="10" | [[Image:20140801 132828.jpg|400px|Today's gel]]<br>
 
| rowspan="10" | [[Image:20140801 132828.jpg|400px|Today's gel]]<br>
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[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 
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| DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) || 3.0 μL
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| DNA(BL09x3, CMV/MV9x3) || 3.0 μL
 
|-
 
|-
 
| 10X buffer || 1.5 μL
 
| 10X buffer || 1.5 μL
 
|-
 
|-
| XbaI || 1.0 μL
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| XbaI || 1.0 μL(or 0 for dephos CMV/MV9)
 
|-
 
|-
| NruI || 1.0 μL
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| NruI || 1.0 μL (or 0 CMV/MV9 and dephos CMV/MV9)
 
|-
 
|-
 
| dH<sub>2</sub>O || 8.5 μL
 
| dH<sub>2</sub>O || 8.5 μL

Latest revision as of 00:09, 27 September 2017


Hayneslab3.gif
Asu logo 3.gif

Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Summary

Ligation Digest and Gel Verification

Reagent Volume

Expected:
BL09 Ligation Colonies 1-3[XbaI/NruI] (Well: 2-4)
Forward insertion: 710, 5921
Reverse insertion: 2141, 4490
No insertion: 710, 4490

CMV/MV9 with [XbaI/NruI] (Well: 5)
710, 4490

CMV/MV9 [XbaI](Well: 6)
5200

Dephos CMV/MV9 (Well: 7)
5200

Today's gel

15 μL/lane; 1% agarose;
Observed lengths:
W2(BL09Lig1): 5000, 7100, 2500;
W3(BL09Lig2): 5000, 7100, 2500;
W4(BL09Lig3): 5000, 7100, 2500;
W5(Empty CM9): 5000, 700;
W6(CM9XbaI): 10000, 20000, 5000.
W7(DephosCM9): 5000
Ladder

DNA(BL09x3, CMV/MV9x3) 3.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL(or 0 for dephos CMV/MV9)
NruI 1.0 μL (or 0 CMV/MV9 and dephos CMV/MV9)
dH2O 8.5 μL
Total 15 μL --> 37°C/ 30 min.