User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/09/25: Difference between revisions
From OpenWetWare
Sean P Corum (talk | contribs) |
Sean P Corum (talk | contribs) |
||
Line 72: | Line 72: | ||
*# Read: 625/655 nM @ gain = 225 w/ settings | *# Read: 625/655 nM @ gain = 225 w/ settings | ||
*# Loop: 2-5 every 3 m for 12 h | *# Loop: 2-5 every 3 m for 12 h | ||
** settings = | ** settings = high lamp energy, endpoint, bottom reading, other settings default | ||
* Transcription maximized around 1 hr after the experiment began. I fine-tuned the measurements, zeroing in on the range of gain = 210-212 as the maximum possible gain where counts < 80,000. I found that the 10 nM plasmid sample broke that threshold at gain = 212, but didn't at gain = 211. | * Transcription maximized around 1 hr after the experiment began. I fine-tuned the measurements, zeroing in on the range of gain = 210-212 as the maximum possible gain where counts < 80,000. I found that the 10 nM plasmid sample broke that threshold at gain = 212, but didn't at gain = 211. | ||
* Therefore, gain = 211 is the maximum possible gain, achieving the best possible fluorescence sensitivity, without breaking the 80,000 count threshold at 10 nM plasmid. | * Therefore, gain = 211 is the maximum possible gain, achieving the best possible fluorescence sensitivity, without breaking the 80,000 count threshold at 10 nM plasmid. | ||
Line 82: | Line 82: | ||
*# Shake: fast double orbital 30s | *# Shake: fast double orbital 30s | ||
*# Loop: 3-5 every 3 m for 12 h | *# Loop: 3-5 every 3 m for 12 h | ||
** settings = | ** settings = high lamp energy, endpoint, bottom reading, other settings default |
Revision as of 15:21, 26 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.
|