Haynes Lab:Notebook/CRISPR Editing/2015/09/08

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09/08/2015 Purified PCR products, set up annealing reactions for 24h time point Cell type drug gRNA time post xfect rep 260 280 260/280 ng/µL ul for 400ng ul ann buff ul water Gal4 dox g034 24 1 0.076 0.042 1.813 76.239 5.25 1.50 8.25 Gal4 dox g034 24 2 0.087 0.048 1.812 87.35 4.58 1.50 8.92 Gal4 dox g034 24 3 0.08 0.043 1.851 80 5.00 1.50 8.50 Gal4 puro g034 24 1 0.085 0.048 1.788 85.425 4.68 1.50 8.82 Gal4 puro g034 24 2 0.05 0.029 1.725 50.174 7.97 1.50 5.53 Gal4 puro g034 24 3 0.078 0.044 1.783 77.729 5.15 1.50 8.35 Ran nested PCR for the remaining samples for the time course experiment. 1:1000 dilution, P149/176, staggered the set up annealing reactions and surveyor for 48h and 72h, ran on BA: Cell type drug gRNA time post xfect rep 260 280 260/280 ng/µL ul for 400ng ul ann buff ul water Gal4 dox g034 48 1 0.087 0.049 1.789 87.049 4.60 1.50 8.90 Gal4 dox g034 48 2 0.089 0.049 1.808 88.656 4.51 1.50 8.99 Gal4 dox g034 48 3 0.08 0.043 1.853 79.666 5.02 1.50 8.48 Gal4 puro g034 48 1 0.08 0.04 1.972 79.795 5.01 1.50 8.49 Gal4 puro g034 48 2 0.086 0.045 1.911 85.877 4.66 1.50 8.84 Gal4 puro g034 48 3 0.069 0.034 2.024 68.808 5.81 1.50 7.69 Cell type drug gRNA time post xfect rep 260 280 260/280 ng/µL ul for 400ng ul ann buff ul water Gal4 puro g034 72 1 0.084 0.044 1.91 84.274 4.75 1.50 8.75 Gal4 puro g034 72 2 0.069 0.036 1.92 69.015 5.80 1.50 7.70 Gal4 puro g034 72 3 0.084 0.044 1.921 83.748 4.78 1.50 8.72 Gal4 dox g034 72 1 0.063 0.032 1.937 62.92 6.36 1.50 7.14 Gal4 dox g034 72 2 0.059 0.031 1.919 59.311 6.74 1.50 6.76 Gal4 dox g034 72 3 0.078 0.041 1.889 78.341 5.11 1.50 8.39