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

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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, 11-14), BL05(Well:2)
Forward insertion: 710, 7010.
Reverse insertion: 3230, 4490.
No insertion: 710, 4490.


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


Today's gel

15 μL/lane; 1% agarose; Ladder

Observed lengths:
W2(BL05Lig1): 5000, 700;
W4(BL01Lig1): 3200, 4500;
W5(BL01Lig2): 5000, 700;
W6(BL01Lig3): 5000, 700;
W7(BL01Lig4): 5000, 700;
W8(BL01Lig5): 5000, 700;
W9(Empty CMV/MV9): 5000, 700;
W11(BL01Lig6): 5000, 700;
W12(BL01Lig7): 5000, 700;
W13(BL01Lig8): 5000, 700;
W14(BL01Lig9): 5000, 700;

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.


One reverse insertion. No forward insertions. Will reattempt ligation.


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