I am trying to do the 3D PCR and realized that the amount of adding DMSO or Betaine influences the outcome of the second nested PCR. Could anyone send me a protocol for the first and second PCR (e.g. amount of DMSO or Betaine, primers, MgSO4,...)
These would be empirical trial and error findings in your case. But for some general parameters, see the 'Additives and enhancers... (starting in the middle of page 44)' section attached book chapter - perhaps there are some helpful bits of info there.