Colony PCR


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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