difference between PCR primers and sequencing primers (View forum version)


Posted 15 January 2011 - 10:19 PM

Is there any differences between primer design rules for PCr and sequencing reactions?
actually I design some primers for sequencing according to my experience on pcr primer design but it didnt work, so is there any special issue in designing sequencing primers?



Posted 20 January 2011 - 07:37 AM

You can often use the primers you used for PCR for sequencing reactions, however some will work much better than others (don't really know why). I would try and when it doesn't work, I'd redesign internal primers.
What is the annealing temperature of your PCR?