User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/09/25: Difference between revisions
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* Cell-free expression 80%: | * Cell-free expression 80%: | ||
** 30 μL extract (12May11) | ** 30 μL extract (12May11) | ||
** | ** 3.1 μL water | ||
** 9 μL 100 μM malachite green (10 μM final) | |||
** 0.9 μL 100 mM Mg-glutamate (1 mM final) | ** 0.9 μL 100 mM Mg-glutamate (1 mM final) | ||
** 2 μL 3 M K-glutamate (66.7 mM final) | ** 2 μL 3 M K-glutamate (66.7 mM final) | ||
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*# 1 μL 50 nM / 3^0 * 20% = 10 nM | *# 1 μL 50 nM / 3^0 * 20% = 10 nM | ||
* Plate reader: | * Plate reader: | ||
*# 29 °C | |||
*# Shake: fast double orbital 30s | |||
*# Read: 625/655 nM @ gain = 175 w/ settings | |||
*# Read: 625/655 nM @ gain = 200 w/ settings | |||
*# Read: 625/655 nM @ gain = 225 w/ settings | |||
*# Loop: 2-5 every 3 m for 12 h | |||
** settings = full plate, high lamp energy, endpoint, bottom reading, other settings default |
Revision as of 12:28, 25 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> |
Characterization B-C: Expression of PHIX174 promoters/UTRs fused to PX-UTR1-deGFP and PX-UTRX-deGFP.
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