Jones Lab:Protocols:PCR

PCR

Add the following to a PCR tube (Master mix if necessary)

1. 10 μL H₂O
2. 2 μL 10x PCR buffer
3. 2 μL 2.5mM dNTPs (each)
4. 2 μL 20mM MgCl₂
5. 1 μL Forward primer (10μM)
6. 1 μL Reverse primer (10μM)
7. 1 μL taq polymerase
8. 1 μL DNA

Mix gently, then setup PCR cycles

1. 94°C for 2 mins
2. 94°C for 30 secs
3. 58°C for 15 secs
4. 72°C for 1 min per Kb of amplicon
5. Go to #2 35 times
6. 72°C for 5 mins
7. 10°C hold