User:Bipedalape/1 July 2008

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<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:
 +
95  3min  
95  3min  
 +
{95  0.5 min
{95  0.5 min
 +
50  0.5 min
50  0.5 min
 +
72  1 min)
72  1 min)
Line 10: Line 26:
72  10 min
72  10 min
 +
4    hold
4    hold
PCR Product of ~300bp (Successful)
PCR Product of ~300bp (Successful)
w/o DMSO
w/o DMSO
 +
 +
<br>Verification of Antibiotic Plates Potency: Colonies were isolated on Chloro, Kana, Amp, and Tet. Successful
<br>Verification of Antibiotic Plates Potency: Colonies were isolated on Chloro, Kana, Amp, and Tet. Successful

Latest revision as of 19:39, 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

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