Haynes Lab:Notebook/Engineering PC-TFs/2014/11/12

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Summary

Restriction Digest/Gel Verification

Reagent Volume
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.


Expected:

BL01
(LCR)Forward insertion: 710, 7010.
Reverse insertion: 3230, 4490.
No insertion: 710, 4490.


CMV/MV9 [XbaI/NruI]
710, 4490


Today's gel

15 μL/lane; 1% agarose; Ladder

Observed lengths:
W2(BL01LCR): 4900;
W3(BL01LCR): 4900;
W4(BL01LCR): 4900;
W5(BL01LCR): 4900;
W6(Empty CMV/MV9):
W7(BL01 Long Transformation): 9000, 2500, 1600;
W8(BL01 Long Transformation): 9000, 2500, 1600;
W9(BL01 Long Transformation): 9000, 2500, 1600;
W10(BL01 Long Transformation): 9000, 2500, 1600;


Retransform LCR on Amp plate to see if colonies form (to see if there is actual plasmid DNA and not some weird genomic DNA artifact). Or PCR with DD123 and DD122.