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

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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
1 Phusion P196 P198 1522
2 Phusion P197 P198 1660
3 Taq 2x P196 P198 1522
4 Taq 2x P197 P198 1660

Expected band sizes:

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

14.09.10 C on Luc SRVYR primer test.png

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.

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 3 4c 2 P192 P195 918

Annealing Temp 57, et 1min