User:Cassandra M Barrett/Notebook/Open Chromatin/2015/09/26

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LCR Transformations and gel results Purpose: to test effectiveness of LCR by transforming E. coli with ligated DNA Methods: Reactions: Reaction 1 Reaction 10 Reaction 11 MV10 uncut MV10 XbaI/MBN/PNK MV10 XbaI/MBN/ not PNK Water Follow standard DH5alpha transformation protocol: Add 50uL of DH5alpha turbo cells to each tube with appropriate sample (2.5uL of each) Incubate on ice for 30min Heat shock at 42C for 35sec Incubate on ice for 2min Add 750uL of SOC media (RT) to each tube Shake 37C for 1hr Plate on LB+Amp (Spin down, remove 200uL of supernatant, resuspend and plate 100uL) Incubate overnight at 37C Run 2uL of reactions 1-11 on gel We see vague larger bands on 10 and 11, probably just reannealed MV10 No sample in rxn 8 tube for some mysterious reason....