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[laska]

Hey
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.

MJL

[phage434]

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.