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

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03/03/2015 Checked concentrations on minipreps from last year of Lonza pMax GFP from Krishna. very low so JW transformed the original along with pMax GFP 6.2 and KAH60. Sample Read# 260 280 260/280 ng/µL pMax GFP 0.026 0.015 1.736 26.294 pMax GFP 0.023 0.013 1.766 23.175 pMax GFP 0.031 0.019 1.639 30.851 Ran PCR products from P151/158 on gel. Only single product for each band. Sequencing: Need to purify. Will purify 25ul and elute into 35 ul. Purification results: Cell type treatment rep 260 280 260/280 ng/µL Luc14 Untreated 1 0.035 0.017 2.006 35.007 Luc14 g046 1 0.032 0.016 2.02 31.691 Luc14 Untreated 2 0.045 0.022 2.034 44.797 Luc14 Untreated 3 0.046 0.024 1.938 46.277 Gal4-EED +dox Untreated 1 0.04 0.019 2.099 40.212 Gal4-EED +dox Untreated 2 0.043 0.023 1.885 42.773 Gal4-EED +dox Untreated 3 0.044 0.023 1.93 43.675 Luc14 g046 2 0.03 0.015 2.086 30.259 Luc14 g046 3 0.031 0.015 2.113 31.368 Gal4-EED +dox g046 1 0.035 0.017 2.042 35.122 Gal4-EED +dox g046 2 0.039 0.019 2.05 38.501 Gal4-EED +dox g046 3 0.041 0.022 1.897 41.336 Luc14 Untreated 1 0.032 0.016 1.949 31.937 Luc14 g046 1 0.03 0.015 1.94 29.645 Sent for sequencing at biodesign with P176 Label Cell type treatment rep PCR primer 1 Luc14 Untreated 1 P176 2 Luc14 Untreated 2 P176 3 Luc14 Untreated 3 P176 4 Gal4-EED+dox Untreated 1 P176 5 Gal4-EED+dox Untreated 2 P176 6 Gal4-EED+dox Untreated 3 P176 7 Luc14 g034 1 P176 8 Luc14 g034 2 P176 9 Luc14 g034 3 P176 10 Gal4-EED+dox g034 1 P176 11 Gal4-EED+dox g034 2 P176 12 Gal4-EED+dox g034 3 P176 13 Luc14 g046 1 P176 14 Luc14 g046 2 P176 15 Luc14 g046 3 P176 16 Gal4-EED+dox g046 1 P176 17 Gal4-EED+dox g046 2 P176 18 Gal4-EED+dox g046 3 P176 Ran PCR with P151/158 on g023, g045, g048 PCR with P197/198 diluted 1:1000 from yesterday. Ran on gel.