Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/04/02
|gBlock (DBN001) & DBN002 PCR Amplification; pSB1A3-GFP Restriction Digest|| Main project page|
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Running out of the gBlock ordered (DBN001), and it's not yet in a plasmid, so I'm making more by PCR amplification.
gBlock PCR run details:
Run is saved as DBN20140402B on the Bio-Rad thermal cycler.
DBN002 PCR Run
Run is saved as DBN20140402A on the Bio-Rad thermal cycler.
pSB1A3-GFP plasmid digestion
Mix and incubate at 37°C for 15 minutes. Then heat-inactivate by incubating at 80°C for 20 minutes.
Gel Electrophoresis of DBN001, DBN002 and pSB1A3-GFP Products
It looks like I ran the gel for too long, so the expected size fragment for the gBlock would have run into the pSB1A3-GFP lane. Will need to run that again. It also looks as though the pSB1A3-GFP digestion was not successful. Will need to re-try with more plasmid, as I used most of it on transfecting the cells.