Haynes Lab:Notebook/CRISPR Editing/2015/05/18

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05/18/2015

Colony PCR of colonies on ligation plates from Saturday. Primers P220/221 will give a 1240bp band if GFP is inserted. will give a 439bp for backbone only. Will be slightly longer if the backbone was only cut once. Used 2x taq MM. NEB suggests an annealing T of 51°C with Taq, will do 55°C to try to avoid lots of off target binding to gDNA.

Testing Neon nucleofector
Resuspension buffer R
GFP control plasmid pMaxGFP6.2