Has anyone tried to increase the Tm of a forward primer with additional random primers at the 3' end? I'm trying with 6N, but it's not working.

I'm trying a gradient PCR to check the best T, previously I have been using 62 as the best option. I would like to increase it to get closer to the Reverse primer, that is 71 degrees. I am using...

14 December 2012 9,080 15 View