Primer-dimer problem - (Aug/24/2004 )
After doing the PCR, when I run the sample on gel I can see the band of my interest, but at the same time I see primer dimer. Could you please suggest how can I reduce primer-dimer formation and also, if I go ahead with such sample for PCR purification...does primer dimer interferes with purification of the product. Any suggestions would be highly appreciated.
Is your product well separated from the primer dimer? If so, then go ahead and do the purification of your product. You may increase the annealing temperature to reduce the primer dimerization. You may also want to use higher percentage gel.
just cut your desired band out from the gel, purify it and you should be fine!?
or do i misunderstand your question?
Thanks guys, for your quick response.
How, if I do hot start PCR to reduce dimerization? Does this work. Pl.let me know.
Thanks a bunch