"
User:Bipedalape/1 July 2008
From 2008.igem.org
(Difference between revisions)
Bipedalape (Talk | contribs) |
Bipedalape (Talk | contribs) |
||
| Line 1: | Line 1: | ||
<br>PCR: MioC (Promoter Region from JM109) | <br>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: | PCR settings: | ||
Revision as of 19:38, 25 October 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
