Haynes Lab:Notebook/CRISPR Editing/2014/09/10

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09/10/2014

Did a test PCR for CRISPR on chromatin samples. Looks great, will start with all samples tomorrow.

Sample PCR mix F primer R primer Band
1PhusionP196P1981522
2PhusionP197P1981660
3Taq 2xP196P1981522
4Taq 2xP197P1981660



Expected band sizes:

1522 for lanes 1 & 3
1660 for lanes 2 & 4




Checked Luc->AmCyan PCR donor I transfected with on a gel. The correct band is there but is faint with other bands so I will do the PCR again and raise the at and reduced the amount of template to try to get rid of other bands.
300p

Ran PCR of PCR 4c 2 (PCR 4c 3), 100μL reaction, 1μL of each primer, .5μL template.

Sample Template F primer R primer Exp band
4c 34c 2P192P195918

Annealing Temp 57, et 1min



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