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

Project name

Main project page Previous entry      Next entry

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 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 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