Posted 11 June 2010 - 05:06 PM
I am trying to introduce a single mutation, and I think my problem may be that the region I am targeting is very GC rich. The oligos I have got are about 75% GC.
I have tried two different annealing temperatures (60 and 55 degrees), 5% DMSO, decreasing the primer concentration and increasing the template concentration without any progress.
Any help would be greatly appreciated. I read somewhere else that trying an annealing temp of 68 might help.
Posted 17 June 2010 - 12:57 PM
Posted 17 June 2010 - 01:01 PM