Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/12/11
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Repeating PCR of pSB1A3_BsmBI fragment.
Template: DBN001_pSB1A3 (close enough) Primers: P140, P141
PCR protocol: basic Phusion, 60°C annealing temp, 90 second extension time, 35 cycles.
No amplification, because no binding site for reverse primer on selected template. Picked up some I13422_pSB1A3 from Wang Lab (pSB1A3 vector with pTet & GFP insert), that should work better. Will repeat attempt on Monday.