User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/08/20: Difference between revisions
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===Hypothesis 2: Gene L is necessary for phage propagation.=== | ===Hypothesis 2: Gene L is necessary for phage propagation.=== | ||
* Here is the quantiluore results from the WP-PCR of 0.1 nM ΦX174 with variable 36-mer primer concentration. ΦX174 1 nM template only control gave 3.34 ng/μL = 0.97 nM. | * Here is the quantiluore results from the WP-PCR of 0.1 nM ΦX174 with variable 36-mer primer concentration. ΦX174 1 nM template only control gave 3.34 ng/μL = 0.97 nM. WAIT: Shouldn't this be 0.1 nM? It seems like I am measuring things to be one order of magnitude too high. | ||
*# 0 nM - 2.82 ng/μL = 0.82 nM = 0.85 fold amplification | *# 0 nM - 2.82 ng/μL = 0.82 nM = 0.85 fold amplification | ||
*# 100 nM - 245 ng/μL = 71 nM = | *# 100 nM - 245 ng/μL = 71 nM = 730 fold amplification | ||
*# 200 nM - 270 ng/μL = 78 nM = | *# 200 nM - 270 ng/μL = 78 nM = 810 fold amplification | ||
*# 500 nM - 303 ng/μL = 88 nM = | *# 500 nM - 303 ng/μL = 88 nM = 910 fold amplification | ||
*# 1000 nM - 320 ng/μL = 93 nM = | *# 1000 nM - 320 ng/μL = 93 nM = 960 fold amplification | ||
* Next step is to optimize primer concentration over a range of 0, 1, 2, 5, and 10 μM (each). I made 10X primer 4 mixes (0, 10, 20, 50, and 100 μM) over this concentration range. 50 μL + 10% WP-PCR reaction. | * Next step is to optimize primer concentration over a range of 0, 1, 2, 5, and 10 μM (each). I made 10X primer 4 mixes (0, 10, 20, 50, and 100 μM) over this concentration range. 50 μL + 10% WP-PCR reaction. |
Revision as of 16:12, 21 August 2012
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Hypothesis 2: Gene L is necessary for phage propagation.
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