Protocols PCR Mutation

Set up two separate primer extension reactions One for each top and bottom primer

Cocktail: 40ul H2O 5ul 10X Pfu Buffer 1ul dNTP (10mM) 1ul Pfu Polymerase 1ul of each Primer (10uM) 1ul Template _____________ 50ul ThermoCycle Program 94*C.......30 sec _____________ 95*C.......30 sec 55*C.......60 sec 68*C.......2 min/kb Run for a total of 4 cycles Combine 25ul from each extension reaction above. Add 1ul Pfu Polymerase. Incubate as above for a total of 18 more cycles.

Protocols