Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/07/22
|Sequence verification around tk-luc insert: PCR||Main project page|
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In order to verify that the luc replacement is successful we will have to sequence the region around the gene before and after using primers specific to the old and new inserts. I previously extracted genomic DNA from Gal4EED-luc HEK293 cell line and will be performing PCR to amplify the regions immediately upstream and downstream of the insert.
Reaction 1 (anneal temp. 54°C):
Reaction 2 (anneal temp. 62°C):
Gel Electrophoresis Results
2nd reaction worked, 1st did not. Will try and repeat using lower temperature and gradient.