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Does PFU creat overhangs - (Nov/04/2008 )

I know that TAQ polymerase tends to create A overhangs when producing PCR products. I was wondering if the same is true for PFU polymerase or if this enzyme creates blunt ends.



no, pfu leaves blunt ends


yes pFU only create blunt end.

Regarding the Taq. it can also add other single base provided that you add specific dNTP and only that single type into the reaction. In other owrds, you could use it to produce single base overhang i.e. T on blunt ended plasmid, sort of like TOPO cloning but cheaper and more time consuming . this plasmid could be used to ligate PCR product amplified using Taq polymerase with normal T4 DNA ligase.