Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/10/30
|Today's project is...||Main project page|
Previous entry Next entry
Phusion gradient PCR.
Reaction volume: 160 µL (split into 8 tubes to be pooled later)
All primers have annealing temperature at 60°C
Thermal cycler program:
Top lanes are ZeoR, bottom lanes are HPK-CFP. Annealing temperature does not appear to be a contributing factor. Must have been use of CG buffer or DMSO.
Next steps: PCR cleanup, maybe gel purification of HPK-CFP fragment (to avoid small fragment), LCR.