SBB10Ntbk-ChrisAnderson

JCAnderson 16:25, 21 March 2010 (EDT)

Gel at left is the A+B+C SOEing reaction and the D2 SOEing reaction. Both look pretty decent. Gel below that is a repeat of R, S, T pcrs. S, the attP21 site didn't amplify. Need some bonafide MG1655 methinks. The digesting and ligation strategy is here.

JCAnderson 20:00, 20 March 2010 (EDT)

Above right goes A,B,C,D,G,H,L, and M pcrs for the big cleanup set attached here. Those were all 55 pcrs. I cut out A,B,C to cogp. Also cut out D to mix with 2 wobbles.

The gel below that is the 2K55 PCRs going E, F, I, J, K, N. F is phiC31 -- it looks a little off, seems small, perhaps the faint slightly larger band is the right product. Hmm. Ahh crap...construction file is wrong, should be:

 PCR AS009-F/AS012-R on A+B+C	(1868 bp, EcoRI/BamHI)

OK, will redo that tomorrow.

JCAnderson 20:31, 22 February 2010 (EST)

Above right is a gel of 1) sbb24 pcr; 2) MW; 3) sbb15 pcr; 4) sbb31
sbb15 and sbb31 are switched, I switched them back. sbb24 looks fine, I zymo'd it, pdt labeled "sbb24 pcr cleanup". Tubes put in box B.