Haynes Lab:Notebook/Engineering PC-TFs/2014/06/02

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Summary

Restriction Digest

Reagent Volume Expected:
Forward insertion: 710, 7010
Reverse insertion: 3230, 4490
No insertion: 710, 4490.
Today's gel
15 μL/lane; 1% agarose; Observed lengths in C1-3, ~3800, ~1950; Ladder
BL01 CMV/MV9 Ligation product colonies 1, 2, 3 respectively 5.0 μL
10X buffer 1.5
XbaI 1.0
NruI 1.0
dH2O 6.5
  15 μL --> 37°C/ 35 min.

BL05 & BL09 Cloning
Followed standard transform procedure.

  • Mixed 50μL thawed DH5α-T cells with 0.5μL BL05 and BL09 plasmid respectively in 10μL dH2O. Incubated for 5 minutes on ice. Plated. Put in incubator.




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