User:Karmella Haynes/Notebook/BioBrick cloning/2010/05/28

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05/28/10 ✓ Minipreps: MV1 (2), MV7 (2) (no methylation) Minipreps > Check with BsaBI/ BstBI digests (methylation sensitive) > Use old preps from DH5α as a control Reagent Volume Expected: 1,2. MV1 (dam-) = 823, 4605 3. MV1 (dam+) = uncut 4,5. MV7 (dam-) = 823, 3919 6. MV7 (dam+) = uncut 15 μL/lane; 1% agarose DNA(plasmid) 2.0 μL 10X buffer 1.5 EcoRI 1.0 PstI 1.0 dH2O 9.5   15 μL --> 60°C (BsaBI)/ 1 hr.; 65°C (BstBI)/ 1 hr. --> Success! C2925 cells produce non-methylated plasmids. Proceed w/ cloning hygro (PCR) into these plasmids.