Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/12/14

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Overview Repeating PCR of pSB1A3, this time with the correct template. Materials/Methods Reagent Stock Volume (µL) GC Buffer 5x 10 dNTP mix 10 mM 1 DMSO - 1.5 Phusion 20 U/µL 0.5 Template 50 ng 0.3 Primer (for) 100 µM 0.2 Primer (rev) 100 µM 0.2 H2O - 36.3 Total 50 Template:I13522_pSB1A3 Primers: P140, P141 PCR protocol: basic Phusion, 60°C annealing temp, 90 second extension time, 35 cycles. Results See rightmost lane. Band visible in the correct size range. Looks like the PCR worked. Performed PCR cleanup on pSB1A3_BsmBI, measured concentration using Take3 plate reader. Final concentration: 31.4 ng/µL