How to get rid of larger non-specific bands appearing in 1st round of pcr just above the expected bands (of approx. 1.5 kb)? The non-specific bands from the 1st pcr are amplified in 2nd round of pcr using nested pair of primer.
Are you asking or just saying us?
I would try to optimize the first rund of PCR. Tips and tricks to do thta can be found here:
I had the same problem before. I advice you to use an adjuvant to improve your PCR effeciency. I used Q solution that comes with hotstar PCR master mix and it gives me a very good amplicon.