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primer orientation - (Nov/21/2005 )

I need help with ordering primers to amplify an ORF. If a gene is in forward (-->) orientation in the genome I order the 5' primer which is the same as the genome sequence and the 3' primer as the reverse complement to the sequence. Right? What if the gene is in the reverse (<--) direction in the genome?


doesn't matter

you will have a piece of double-stranded DNA after your PCR reaction

what you do with the fragment later will be determined by the gene's orientation; you will still have the same piece of DNA, right?