Haynes Lab:Notebook/Engineering PC-TFs/2014/09/01

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(Summary)
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==Summary==
==Summary==
-
*  
+
* Split cells.
 +
* Restriction digest/gel verification of PcTF CMV/MV9 ligation.
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----
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''''Restriction Digest/Gel Verification''''
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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><b>Expected:</b></u>
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<br>
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<br>BL01(Well: 4-8, 10-13), BL05(Well:14) <br>
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Forward insertion: 710, 7010. <br>
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Reverse insertion: 3230, 4490. <br>
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No insertion: 710, 4490. <br><br>
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 +
 
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<u>CMV/MV9 [XbaI/NruI] (Well: 9)</u> <br>
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710, 4490<br><br>
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| rowspan="10" | [[IMAGE URL HERE|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(BL01Lig1): 5000, 700; <br>
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W3(BL05Lig1): 5000, 700; <br>
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W4(BL05Lig2): 5000, 700; <br>
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W5(BL05Lig3): 5000, 700; <br>
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W6(BL05Lig4): 5000, 700; <br>
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W7(Empty CMV/MV9): 5000, 700; <br>
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W8(BL09Lig1): 5000, 700; <br>
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W9(BL09Lig2): 7000, 700; <br>
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W10(BL09Lig3): 5000, 700; <br>
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W11(BL09Lig4): 5000, 700; <br>
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W12(BL09Lig5): 7000, 700;<br>
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W13(BL09Lig6): 5000, 700; <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) || 2.0 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 || 9.5 μL
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|-
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| Total || 15 μL --> 37°C/ 30 min.
 +
|}

Revision as of 18:46, 1 September 2014



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Summary

  • Split cells.
  • Restriction digest/gel verification of PcTF CMV/MV9 ligation.

'Restriction Digest/Gel Verification'

Reagent Volume

Expected:

BL01(Well: 4-8, 10-13), BL05(Well:14)
Forward insertion: 710, 7010.
Reverse insertion: 3230, 4490.
No insertion: 710, 4490.


CMV/MV9 [XbaI/NruI] (Well: 9)
710, 4490


400px|Today's gel

15 μL/lane; 1% agarose;

Observed lengths:
W2(BL01Lig1): 5000, 700;
W3(BL05Lig1): 5000, 700;
W4(BL05Lig2): 5000, 700;
W5(BL05Lig3): 5000, 700;
W6(BL05Lig4): 5000, 700;
W7(Empty CMV/MV9): 5000, 700;
W8(BL09Lig1): 5000, 700;
W9(BL09Lig2): 7000, 700;
W10(BL09Lig3): 5000, 700;
W11(BL09Lig4): 5000, 700;
W12(BL09Lig5): 7000, 700;
W13(BL09Lig6): 5000, 700;
Ladder

DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) 2.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
NruI 1.0 μL
dH2O 9.5 μL
Total 15 μL --> 37°C/ 30 min.



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