Talk:PCR Amplification of DNA
From Bridges Lab Protocols
from [Damn Good Technician] suggestions:
What polymerase do I use, you ask? Why, it's KOD polymerase, from EMD Biosciences. For all I know, it's the polymerase everyone uses. If it isn't, let me tell you the three basic reasons why you ought to use it. Reason 1: it has remarkable fidelity. I've never had a PCR-introduced base pair change in all the cloning I've done with it, and I've done quite a lot. Reason 2: it has blazing speed. My standard PCR extension times for a 3kb amplicon are around 30s, with annealing times of around 3-4s. If you don't pour your agarose gel to run your PCRs before you set up your PCR, it won't have solidified by the time your PCRs are done - it's that fast. Reason 3: it's very good with GC rich templates (specifically genomic templates), and also very long templates (also occasionally genomic templates)..
I'll order in some KOD polymerase and try a 30s extension with a 3-4s annealing