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

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Summary

  • Concentration reading for gel purified PcTFs, ligation product minipreps.
  • Gel verification of ligation attempt 3.

Concentration Readings

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Gel Verification

Reagent Volume

Expected:

BL01(Well: 2), BL05(Well:3-6)
Forward insertion: 710, 7010.
Reverse insertion: 3230, 4490.
No insertion: 710, 4490.


CMV/MV9 [XbaI/NruI] (Well: 7)
710, 4490


BL09 Ligation Colonies 1-6[XbaI/NruI] (Well: 8-13)
Forward insertion: 710, 5921
Reverse insertion: 2141, 4490
No insertion: 710, 4490

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.


BL09Lig2&5 sent in for sequencing.


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