previous I optimised my designed primers without DMSO, but now my subject samples getting problem so I added DMSO, getting good intensity bands along with non specific bands.
So now again optimized with DMSO getting good intensity bands along with non specific bands.
Paul Rutland
try running a gradient pcr or just increasing the annealing temperature by 1C at a time . If this fails select a good amplification condition and try lowering both the primer concentration and the MG concentration. Try also using a hot start enzyme to minimise mis priming
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