Does anyone having experience with using Phusion Hi-fidelity and KOD hot-start Hi-fidelity DNA polymerases and what is the difference between these two. while i have used phusion for amplifying my genes for cloning I don't have any idea bout the other. so if someone have experience in using both could tell me the difference?
#Cloning #DNA #Polymerase #phusion #KOD #jumpstart #genes
I haven't noticed a huge difference overall. KOD is a bit faster, so it will save you time on your PCR reactions. Both are high fidelity, so you don't have to worry much about random mutations.
As you can tell from the name, the KOD "hot-start" shows activity only after initial heating, reducing side reactions while pipeting your reactions.
The main difference is maybe that Phusion polymerase has the highest proofreading-activity of all polymerases that I have worked with, incl. Pfu which has a 2-3 fold higher error rate, still much lower than Taq but not as good as Phusion (depending on manufacturer, you will see different numbers) but these are numbers that we got years ago when we quantified the error rates of different DNA polymerases. KOD seems to have a similar error rate as Pfu (according to SigmaAldrich).
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