Haynes Lab:Notebook/CRISPR Editing/2015/08/27

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08/27/2015 Collected g034 transfections at 48 hours. Flow and gDNA extraction, three wells for g4+p and three wells for g4+d Set up annealing reaction for Surveyor of Luc14 dilution of g034 treated cells collected 7/8/15. Cell type drug ug g034 rep ul for 400ng ul buffer ul water for 15ul Luc14 - 1 1 6.13 1.50 7.37 Luc14 - 1 2 6.03 1.50 7.47 Luc14 - 1 3 6.11 1.50 7.39 Luc14 - 0.1 1 5.98 1.50 7.52 Luc14 - 0.1 2 6.17 1.50 7.33 Luc14 - 0.1 3 6.60 1.50 6.90 Luc14 - 0.01 1 5.87 1.50 7.63 Luc14 - 0.01 2 6.08 1.50 7.42 Luc14 - 0.01 3 6.87 1.50 6.63 Some PCR purifications from that same dilution experiment didn't have high concentrations so redid PCR with P163/215 Gal4-EED + dox 0.1ul g034 rep 3 Gal4-EED + dox 0.01ul g034 rep 1 Set up annealing reaction for Surveyor of Luc14 singlet from initial screen of gRNAs around the end of 2014 Luc14 gRNA 260 280 260/280 ng/µL ng/µL average ul for 400ng ul ann buff ul H2O Luc14 g029 0.073 0.039 1.889 76.357 73.096 74.73 5.35 1.50 8.15 Luc14 g030 0.043 0.023 1.889 43.492 43.386 43.44 9.21 1.50 4.29 Luc14 g033 0.084 0.045 1.837 80.736 83.56 82.15 4.87 1.50 8.63 Luc14 g036 0.082 0.044 1.88 82.825 82.088 82.46 4.85 1.50 8.65 Luc14 g040 0.065 0.036 1.794 63.731 64.87 64.30 6.22 1.50 7.28 Luc14 g041 0.055 0.029 1.888 53.011 54.684 53.85 7.43 1.50 6.07 Luc14 g042 0.045 0.024 1.885 47.205 45.157 46.18 8.66 1.50 4.84 Luc14 g043 0.044 0.023 1.88 45.143 43.898 44.52 8.98 1.50 4.52