Haynes Lab:Notebook/CRISPR Editing/2016/06/02

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06/02/2016 Ran g055 nested PCR again, used 05ul of phusion from the wang lab, 32 cycles. ran on gel, bright bands, no band in neg ctrl lane. column purified, set up annealing reaction Cell type gRNA rep 260 280 260/280 ng/µL ul for 200ng ul ann buff ul water Luc14 g055 1 0.098 0.052 1.886 98.084 4.08 1.50 9.42 Luc14 g055 2 0.094 0.049 1.902 94.086 4.25 1.50 9.25 Luc14 g055 3 0.096 0.051 1.884 96.014 4.17 1.50 9.33 ran annealing reaction of g055 and g054. Plate 6 dilution curve for three primer sets. used sheared Luc14 genomic DNA possible plate 7 L 25 H pset 3 L/G 31 F/I/input set 3 Primer set 3 master mix ' 1 well 85 2x SYBR mix 7.5 637.5 750nmol F/R primer mix 3 255 PCR water 2.5 212.5 total 13 1105