during my project, in seminested PCR, appeared the two non specific bands with the one specific band, one of the non specific band is the ame size of first PCR reaction,please, any one can help me, how I can overcome this problem
I think that one of the 'non-specific' bands (which has the same size as the first PCR product) can be just the template of the first PCR product you added as template for the second PCR reaction. Reduce the amount of the template (first PCR Product) may make this 'non-specific' band disappear.