Posted 14 June 2011 - 11:17 AM
Does anyone know of good alternative buffers for use in PCR or strand displacement? Currently I'm using the New England Biolabs thermopol buffer, which is Tris, Triton X-100, MgSO4, KCl, and (NH4)2SO4. But the next step involves a reaction that's inhibited by Tris. So I'm looking into alternative buffers. I've tried desalting and transferring to a PBS buffer (with and without MgSO4), and it didn't work so well. I know the second reaction (a CuAAC) is compatible with PBS. So currently I'm left wondering if I can switch to a different buffer in the first place without disrupting the polymerase reaction. I'm using BST polymerase to convert ssDNA to dsDNA while introducing an artificial nucleotide in place of dTTP. It'd be nice if I could simply switch to a PBS buffer and not worry about desalting.