pfu ultra and single stranded cDNA - (Jun/28/2007 )
A question: Could it be a problem to use pfu ultra with proofreading activity together with single stranded cDNA. Can the enzyme cut the cDNA into pieces making the PCR not work?
should be no problem. if you think about it, you are always amplifying a single strand with pcr (think "denaturing step").
I have used that enzyme to amplify cDNA, just added a part of my RT reaction and that's it. Stratagene recommends longer elongation times for amplifying cDNA, but that's the only difference (don't know why they recommend longer times, and I used normal elongation times).
You are right. I did not think about that!
When I used it with cDNA made in two different ways, with random hexamers and with oligo-dT only the random hexamer cDNA worked.
No problem with pfu for cDNA pcr. just elongation at 68°C instead of 72°C.
72 work but 68 is more efficient.
