15 replies to this topic

[laurequillo]

HomeBrew, on Oct 28 2009, 08:49 PM, said:

It's difficult to be sure what stretch constitutes your matching bases (for example, I count 34 bases after your HindIII site in your reverse primer, but you say there are 31 matching bases)...

However, I think you still have way too many, and your Tm's are too high.

You are right, there are 34.

So, still too long.    

I will follow your advice and go down to 21-25 bases and Tm 60ºC, and I will do a standard pcr

98º 30sec

98º 10sec
55º 20sec
72º 3-4min
    35 cycles

72° 10min