Silly Question about RT-PCR/Primer design - (Oct/25/2006 )

When you design RT-PCR primers are you designing them to target the RNA or the cDNA ? thanks

cDNA

that is the "RT"...you are pcr'ing Reverse Transcribed DNA, or cDNA

Hi,

You design RT-PCR primers targetting the positive strand (or sense strand) of cDNA, which have the same sequence as the RNA, except that the U is T.

So, you should be able to locate the start and stop codon on your template. Not to worry, all nucleotide sequence in Genbank is on the sense strand.

Bests