Haynes Lab:Notebook/CRISPR Editing/2015/01/07

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01/07/2015 Set up 2-50μL PCR reactions for untreated Luc14 gDNA and a subset of treated Luc14. sample Untreated 23 25 29 30 32 33 34 36 39 43 44 45 46 Mastermix 1 ' ' 1rxn 28.3rxns H20 30 849 HF 5X MasterMix 10 283 dNTPs 1 28.3 FP 2.5 70.75 RP 2.5 70.75 46 1301.8 Mastermix 1 1rxn 2.1rxns MM1 46 96.6 Template 2 4.2 DMSO 1.5 3.15 Phusion 0.5 1.05 50 205 Transfer 50μL of MM2 into each of 4-PCR tubes. PCR 98°C for 3 min 36 cycles 98°C for 10s 66°C for 30s 72°C for 60s 72°C for 10 min 4°C forever Sample g039 may have too much gDNA, samples g045 and g046 were in non-autoclaved tubes, sample g046 did not have 50ul for each tube