Team:Exeter/lab book/proto/7

Make-up a 50µL total solution in a PCR tube of:

• 5µL of 10ng/µL genomic DNA template

• 1µL dNTPs

• 1µL Forward Primer (@100pmol/µL)

• 1µL Reverse Primer (@100pmol/µL)

• 10µL High-Fidelity Phusion 5x Buffer

• 1µL High-Fidelity Phusion polymerase

• 31µL MilliQ H2O

Run a two-step PCR experiment using the following settings:

• Stage 1: 98oC for 30 seconds for 1 cycle

• Stage 2: 98oC for 10 seconds followed by 72oC for 20 seconds for 30 cycles

• Stage 3: 72oC for 5 minutes for 1 cycle