Haynes Lab:Notebook/Jan/2016/06/15

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Genomic DNA Extraction, PCR, Agarose Gel Verification, Freezer Inventory Finished extracting genomic DNA from 4-1, 4-2, 4-3, 6-1, 6-2, 6-5, 6-6. PCR: Candidate cell lines (3 primers + 1 error = 4) Reagent Vol Mix* (x #) Expected: 1. 4-x, P1 = 1145 2. 4-x, P3 = 1046 3. 4-x, P6 = 1139 4. 6-x, P1 = 1146 5. 6-x, P3 = 1046 6. 6-x, P6 = 1146 15 μL/lane; 1% agarose; Ladder genomic OR plasmid DNA 1.0 / 0.5 4.0 / 2.0 10 μM forward primer 1.0 --- 10 μM reverse primer 1.0 --- 2x GoTaq green 10.0 40.0 dH2O 7.0 / 7.5 28.0 / 30   20.0 μL 72.0 μL Note: *Prep one Mix per template For each template, aliquot 18.0 master mix into 3 tubes Thermal cycler: Labnet - GOTAQ 95°C 3 min. 30x[95°C, 30 sec; 57°C 30 sec; 72°C 30 sec] 72°C 3 min. 4°C, ∞ Conclusions On the top, left to right: 4-1, 4-2, 4-3, P1, 3, and 6 for each. On the bottom, left to right: 6-1, 6-2, 6-5, 6-6, P1, 3, and 6 for each. -150 Freezer Inventory, Box B6: No 4-1 vials (one active T75 flask) No 4-2 vials (one active T75 flask) One 4-3 vial One 4-4 vial Three 4-5 vials Two 4-6 vials No 4-7 vials (No active flasks, gone forever) No 4-8 vials (No active flasks, gone forever) Two 4-9 vials No 6-1 vials (one active T75 flask) One 6-2 vial Three 6-3 vials No 6-4 vials (No active flasks, gone forever) One 6-5 vial One 6-6 vial One 6-7 vial Two U2-OS wild-type vials