Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/10/28
|Phusion PCR of parts for LCR|| Main project page|
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We're using the Phusion polymerase to create excess template in anticipation of performing ligase-cycling reaction (LCR) in the near future. This will allow us to incorporate a zeocin resistance (ZeoR) gene upstream of the HPK-CFP fluorescent reporter.
Reaction volume: 100 µL (split into 4 tubes to be pooled later)
All primers have annealing temperature at 60°C
Thermal cycler program: