Haynes Lab:Notebook/CRISPR Editing/2015/03/27

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03/27/2015 Ran on gel, purified, and set up digests of nested PCR products from yesterday for g034, unt and g048 Cell type Treatment rep F primer R primer 260 280 260/280 ng/µL ul for 200ng ul for 400ng ul water ul buffer ul enzyme Luc14 g034 1 P163 P215 0.054 0.029 1.834 54.054 7.40 9.60 2 1 Luc14 g034 2 P163 P215 0.053 0.028 1.869 52.537 7.61 9.39 2 1 Luc14 g034 3 P163 P215 0.056 0.03 1.835 55.65 7.19 9.81 2 1 Gal4-EED +dox g034 1 P163 P215 0.055 0.031 1.8 54.901 7.29 9.71 2 1 Gal4-EED +dox g034 2 P163 P215 0.059 0.032 1.851 59.365 6.74 10.26 2 1 Gal4-EED +dox g034 3 P163 P215 0.052 0.027 1.882 51.558 7.76 9.24 2 1 Luc14 Unt 1 P163 P215 0.057 0.032 1.817 57.356 6.97 10.03 2 1 Luc14 Unt 2 P163 P215 0.063 0.034 1.872 63.42 6.31 10.69 2 1 Luc14 Unt 3 P163 P215 0.052 0.027 1.889 51.547 7.76 9.24 2 1 Gal4-EED +dox Unt 1 P163 P215 0.046 0.025 1.831 45.566 8.78 8.22 2 1 Gal4-EED +dox Unt 2 P163 P215 0.049 0.028 1.719 48.879 8.18 8.82 2 1 Gal4-EED +dox Unt 3 P163 P215 0.048 0.027 1.789 47.546 8.41 8.59 2 1 Luc14 g048 1 P213 P214 0.044 0.024 1.805 43.885 4.56 12.44 2 1 Luc14 g048 2 P213 P214 0.043 0.023 1.886 42.798 4.67 12.33 2 1 Luc14 g048 3 P213 P214 0.036 0.02 1.837 36.491 5.48 11.52 2 1 Gal4-EED +dox g048 1 P213 P214 0.033 0.019 1.772 33.381 5.99 11.01 2 1 Gal4-EED +dox g048 2 P213 P214 0.042 0.025 1.707 42.341 4.72 12.28 2 1 Gal4-EED +dox g048 3 P213 P214 0.036 0.021 1.735 36.042 5.55 11.45 2 1 gDNA extraction Cell type Plasmid 1 Plasmid 2 260 280 260/280 ng/µL ul for 200ng ul water to 4 Luc14 pMax GFP 6.2 g034 0.083 0.04 2.052 82.601 2.42 1.58 Luc14 KAH228 pMax GFP 6.2 0.091 0.046 1.993 91.44 2.19 1.81 Luc14 KAH228 g034 0.13 0.067 1.934 129.826 1.54 2.46 Gal4-EED pMax GFP 6.2 g034 0.134 0.069 1.96 134.494 1.49 2.51 Gal4-EED KAH228 pMax GFP 6.2 0.15 0.077 1.95 150.249 1.33 2.67 Gal4-EED KAH228 g034 0.13 0.069 1.886 130.094 1.54 2.46 phusion PCR with P197/198