Haynes Lab:Notebook/Jan/2015/04/10

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04/10/15 Minipreps - BD004/MV2 (6x 5mL cultures) Minipreps - BD004/MV2 ---Karmella 20:52, 10 April 2015 (EDT): Sigma GenElute kit. Elute & back-elute w/ 70 μL elution sln. BD004/MV2 a BD004/MV2 b BD004/MV2 c BD004/MV2 d BD004/MV2 e BD004/MV2 f Restriction Digest check (Fermentas FD) Cut with ApaLI (E/S insertion got rid of BB RFC23 cut sites) Reagent Volume Expected: * BD004/MV2 = 7108, 1243, 288 * MV2 only = 2998, 1243, 288 * BD004/V0120 = 5659, 1246, 497 30 μL/lane, 1% agarose; Ladder DNA (plasmid) 3.0 μL 10x buffer 1.5 ApaLI 1.0 dH2O 9.5   15 μL --> 37°C/ ~10 min. Conclusions Success, and very lucky! Clone "a" is the desired plasmid. Keep clone a and discard other minipreps. Mark the streak library accordingly. Miniprep conc.'s Sample OD260 260/280 ng/μL 1. BD004/MV2 a 0.400 1.90 399.66 2. BD004/MV2 b 0.344 1.90 344.33 3. BD004/MV2 c 0.271 1.88 270.79 4. BD004/MV2 d 0.292 1.89 292.47 5. BD004/MV2 e 0.279 1.90 279.28 6. BD004/MV2 f 0.221 1.88 220.53