User:Meng Xiao He/Notebook/fall08/2008/12/05

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new candidate cco PCR

Phusion, cco Xover

First 11 samples are from 11/30 KO

12-20 and (last 2 are Gm sensitive colony I and non sensitive control colony J from 11/28) did not have bands

Ladder is 1kb

2008-12-05 pcr1.jpg


Remainder of each Rx run again, with extension time increased to 2:30 (to see if WT cco region will PCR properly)> essentially the same results.

What are these?

2008-12-05 22+22 long pcr.jpg

(same done with old cco/cbb/Duo batch and got no fragments)

new conjugation

  • 500 ul MR-1
  • 1mL Duo, 750ul cco, 500ul cbb in 14mL MgSO4> plated on LB+DAP