Fast vs Standard PCR - (Aug/23/2008 )
I wonder what are the key changes that allow a standard PCR to be shortened? And has anyone have experience is transferring protocols from standard to fast PCR systems? What are the success rate?
Which fast system are you using? The lightcycler says you can use the same annealing temp. Set it at 0 seconds. As for the extension time, unless the instrument manual gives you a number, I'd use 30 seconds for most amplicons under 1 kb, then whatever the extension rate is for your polymerase of choice.