User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/07

Pc-TF Genomics

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09/07/14 Minipreps & digests Minipreps Used Sigma kit (ran out of lysis buffer) Resuspended 4 mL cell culture (pelleted) in 400 μL resuspension buffer Lysed with 60 μL Zymo 7x blue buffer Followed rest of Sigma protocol Digests > Check with KpnI digests (same as 9/14/10, Harvard) Reagent Volume Expected: Backbone = 7489 Empty vector frag. = 913 1. KAH160/KAH015-1,2,3 = 2060 2. KAH165/KAH015 = --- 3. KAH170/KAH015-1,2,3 = 1868 15 μL/lane; 1% agarose DNA(plasmid) 1.0 μL 10X buffer 1.5 KpnI 1.0 dH2O 11.5   15 μL --> 37°C/ ~15 min.         --> Reaction conditions