PCR of ssDNA - (Nov/11/2008 )
Hello all!
I am trying to amplify a fragment of 2.5 kb ssDNA but i am having lots of problems. If I amplify a 600 pb I obtain the correct band ( i am using both primers), but when i try to obtain the longer fragment i never manage to obtain it. I have tried with DMSO 2%, longer cycles.....may be it is too much for a ssDNA amplification?. I haven“t done a PCR on a ssDNA before. I am using both primers.
Thank you for your help!
-paramyosin-
What is the source of your ssDNA? Is it cDNA (reverse transcribed and treated with RNase) of how did you get it?
-Trof-
No, it is a ssDNA virus isolated from a sera. I want to amplify part of its genome.
-paramyosin-