User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/09/24: Difference between revisions
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* Ah ha! Since the 1 μM primer mix was diluted from the 10 μM primer mix, it is the case that overly concentrated primers inhibit PCR. | * Ah ha! Since the 1 μM primer mix was diluted from the 10 μM primer mix, it is the case that overly concentrated primers inhibit PCR. | ||
* To optimize this: | * To optimize this: | ||
* | * 0.45 mL 90% ΦX174 WP-PCR Mix (reusable) | ||
** | ** 370 μL H<sub>2</sub>O | ||
** | ** 50 μL 10X reaction buffer | ||
** | ** 15 μL 3.2 nM ΦX174 template (~ 0.1 nM final) | ||
** | ** 5 μL 25 mM dNTPs mix | ||
** | ** 10 μL PfuUltra I DNA polymerase | ||
* Aliquot 5 × | * Aliquot 5 × 4.5 μL: | ||
*# | *# 0.5 μL water = 0 μM primer 4 | ||
*# | *# 0.5 μL 12.5 μM primer mix = 1.25 μM primer 4 | ||
*# | *# 0.5 μL 25 μM primer mix = 2.5 μM primer 4 | ||
*# | *# 0.5 μL 50 μM primer mix = 5 μM primer 4 | ||
*# | *# 0.5 μL 100 μM primer mix = 10 μM primer 4 | ||
* Cycling parameters: | * Cycling parameters: |
Revision as of 16:08, 24 September 2012
PHIX174 Cell Free Expression | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
Hypothesis 2: Gene L is necessary for phage propagation.
Characterization B-C: Expression of PHIX174 promoters/UTRs fused to PX-UTR1-deGFP and PX-UTRX-deGFP.
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