Conventional PCR - (Apr/23/2012 )
I do conventional PCR for HIV single genome sequencing but for four months now my bands form in the wells. My target is 2.4kb and i have been troubleshooting to find out cause but it is not yet clear. My protocol involves a separate cDNA synthesis step. Could any one be facing the same problem or have ant possible solutions to this? Thanks...Kabogere
It could be the result of over-amplification,i.e. too much PCR product has been built (and subsequently loaded on the gel).
Here's a thread where someone has described a similar problem:
Also, this link might help: http://palumbi.stanford.edu/SimpleFoolsMaster.pdf
see p.12 "Bright bands in well of gel"