Protocol Online logo
Top : Forum Archives: : Real-Time PCR

RT-PCR - not qPCR! urgent please - how to choose primers (Oct/06/2006 )

My question:
When I pick the primers for the RT-PCR, should they match the unspliced mRNA or the CDS (the spliced variant that will be translated and starts with the ATG)???

-Jou-

It depends upon if the cDNA was synthetized from mature mRNAs or total mRNAs. If you used poly-A primers for the cDNA, the cds match should work. Good luck!

-aleruiz-

primerbank may be useful too.

-fred_33-

thank you for the advice!

-Jou-