Jessica Karen Wong/Notebook/2007-7-13

I2057

 * Colony PCRed 12 10ul reactions w/ VF and VR, Taq and 1:30 ext time
 * Ran gel (lad, 1-12, 1x DNA, 3x dilution DNA, water, lad)
 * Got lots of too small bands

Sequencing

 * 1) E0240-1AK3 for E0040VR
 * 2) E0240-1AK3 for E0040VF
 * 3) T9002-3K3 for F2620 aka C62 VF
 * 4) T9002-3K3 for C62VR

E0240

 * Redid gradient BB PCR with 3x dilution and pos control P1010-1AK3 at 52.7

I2055

 * Redigested I2055 w/ pcr'ed in promoter w/ mfe and nsi
 * used 4ul dna and buffer 2
 * ligated I2055 M/N (not sure if contains promoter) and 3K3
 * Transformed 3ul of ligation and plated

Ordered new I2056 primers to PCR in Promoter
 * I2056_R0040_F 8mer.Mfe1.promoter+mix.part of RFP
 * CTTAGTAG CAATTG tccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactag ATGGCTTCCTCCGAAGACG (54.0)

Did an overnight colony PCR of
 * T9002 1AK3 (9 samples)
 * T9002 3K3 (16 samples)
 * E0240 3K3 (20 samples)
 * I2055 1AK3 (20 samples)