Haynes Lab:Notebook/Engineering PC-TFs/2014/08/01

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(Summary)
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==Summary==
==Summary==
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'''Ligation Digest and Gel Verification (2)'''
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<br>
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{| {{table}} border="1" cellspacing="5"
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|- valign="top"
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| bgcolor=#cfcfcf | Reagent
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| bgcolor=#cfcfcf | Volume
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| rowspan="9" |
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<u>Expected:</u>
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<br>
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BL09 Ligation Colonies 1-3[XbaI/NruI] (Well: 2-4)<br>
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Forward insertion: 710, 5921 <br>
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Reverse insertion: 2141, 4490 <br>
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No insertion: 710, 4490<br> <br>
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CMV/MV9 with [XbaI/NruI] (Well: 5) <br>
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710, 4490<br><br>
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CMV/MV9 [XbaI](Well: 6) <br>
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5200<br><br>
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Dephos CMV/MV9 (Well: 7) <br>
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5200
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| rowspan="10" | [[Image:20140801 132828.jpg|400px|Today's gel]]<br>
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15 μL/lane; 1% agarose;<br>
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<u>Observed lengths:</u> <br>
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W2(BL09Lig1): 5000, 7100, 2500; <br>
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W3(BL09Lig2): 5000, 7100, 2500; <br>
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W4(BL09Lig3): 5000, 7100, 2500; <br>
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W5(Empty CM9): 5000, 700; <br>
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W6(CM9XbaI): 10000, 20000, 5000.<br>
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W7(DephosCM9): 5000 <br>
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[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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|-
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| DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) || 3.0 μL
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|-
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| 10X buffer || 1.5 μL
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|-
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| XbaI || 1.0 μL
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|-
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| NruI || 1.0 μL
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|-
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| dH<sub>2</sub>O || 8.5 μL
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|-
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| Total || 15 μL --> 37°C/ 30 min.
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|}
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<br>
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__NOTOC__
__NOTOC__

Revision as of 15:53, 3 August 2014



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Summary

Ligation Digest and Gel Verification (2)

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(BL01x2, BL05x1, BL09x2, CMV/MV9x1) 3.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
NruI 1.0 μL
dH2O 8.5 μL
Total 15 μL --> 37°C/ 30 min.




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