User:Bipedalape/1 July 2008

PCR: MioC (Promoter Region from JM109)

Primers:

Reverse - Site Directed Mutation to Produce Spe1 Restriction Site

5' AAC TGC AGC GGC CGC TAC TAG TAA TTG TAC GCT GTG AAC GC 3'

Forward -

5' CGG AAT TCG CGG CCG CTT CTA GAG GCC AAA GAC TAC CCT TCC 3'

PCR settings:

95 3min

{95 0.5 min

50 0.5 min

72  1 min)

(Repeated X 30)

72  10 min

4   hold

PCR Product of ~300bp (Successful) w/o DMSO

Verification of Antibiotic Plates Potency: Colonies were isolated on Chloro, Kana, Amp, and Tet. Successful