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Colony PCR 1. Taq 10X buffer (each tube) a.36 ul H2O b.5 ul buffer thermopole c.1 ul DNTp d.1 ul primer 1 (Brk primer) e.1 ul primer 2 (Brk primer) 2. Add colonies (another set of tubes) a.6 ul H20 b.Colony (pipette up and down) 3. Add 5 ul of colony solution to buffer tube (save the 1 extra ul) 4. Add 1 ul of Taq polymerase to buffer tube 5. Thermocycler – login and use colony PCR protocol a.Change 72 degrees extension time i.Shoot for 0:15 sec longer than necessary ii.Based on the # of base pairs, PCR does 1 kb per minute

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