Hii everyone.
I am trying to amplify a gene specific fragment in wheat genome.Initially i used to get a very good amplicon as a PCR product but after some time , i started observing only a bright smear as a product.I tried all the possible modifications (like gradient, touchdown PCR, modifications in PCR mastermix, etc.) but in vain. Even i re synthesized my primers and also checked my DNA quality and quantity, and it's fine.I also used the same DNA with other primers and i am getting a good result.Can anyone extend the possible solutions to this problem?The below is the gel image of my previous and latest PCR amplification.