Does it help to add antisense primer first for bisulfite PCR? - (Aug/22/2005 )
Because after bisulfite conversion there is no C for sence primer which has G to anneal with. If you do antisense primer first, you can generate new C for G in the sense primer to basepair with.
Does any one have any idea?
you have to design strand specific primers to overcome this issue.
I think it makes sense adding antisense primer first because at the very begining of the amplification, there is no binding site for sense primer. If antisense is added first, after one cycle of amplification, sense primer is followed, in this way, primer dimer can be reduced or avoided.