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User:Bipedalape/1 July 2008
From 2008.igem.org
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
