PCR help minus strand - Primer design (Mar/03/2011 )
I'm new in this field. I would like to detect expression of the X gene in cells, both DNA and RNA level. The gene is on (minus) (-) strand. Shell I change the original sequence somehow when designing primers. Thank you. I'm simply not sure if its necessary to do it, minus strand hmm.
Only you know what is the minus strand. DNA has no notion of orientation (only the transcription and translation are directional). The enzymes in a PCR reaction are insensitive to the orientation -- each dsDNA molecule has one 5' and one 3' base at each end, because the DNA pairs in an anti-parallel fashion.