For a proof of principle experiment I want to perform DNA synthesis on a immobilized ssDNA (~50 bp). I was looking for PCR Polymerases (Taq, etc.) but all of them are of course optimized for high temperatures. Saddly my instrumental set-up has no temperature control and I can only perform the DNA synthesis at room temperature. Can anyone give me an advice for what kind of DNA polymerase I should look?
Thanks a lot for the information, looks like the solution I was looking for.
Theo is right. Use klenow fragment for room temperature. Interestingly, you can also run a parallel experiment with conventional Taq polymerase (optimized for PCR). You may get an amplification, although at a lower amount. I dont know how much of amplification you need, but you can just run an expt and see. I have got quite good results using such Taq for reaction at 25-37 degrees in my experiments.
Good Luck
SD
I agree also with Theo, klenow is good. I expect all polymerases to work at room temperature but the further from the optimal temperature you use them the more error misincorporations there will be . If you held the tube in your hand the temperature would be 37c which would be good also for klenow or just leave it in a water bath not a pcr machine
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