Haynes Lab:Notebook/CRISPR Editing/2015/01/30

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01/30/2015 Prepped KAH126 plasmids for sequencing from the mini preps we did on January 28, 2015 (Wed). We each prepped two tubes. Each contained 7μL of water with 2μL of plasmid template and 1μL of either DD122 primer or DD123 primer. The primers were obtained from Cameron. Picked 6 colonies from one of the g034 topo plates into 2mL LB+amp. Let grow overnight. Added new media to Cameron's cells. Used media+puro from a long time ago, not sure if the puro is still good. Did a test PCR reaction with Untreated Luc14 gDNA and phusion. Only 30 cycles and 6 different annealing temps: 66, 67, 68, 69, 70, 72. Want to see if I can reduce background bands but still get enough product. Put at -20.