GAPDH primer sequences for mRNA level in HeLa cells - (Jul/17/2007 )

Hi everyone,

Can someone share the sequences of a pair of primers that has been SUCCESSFULLY employed for the determination of the GAPDH (glyceraldehyde-3-phosphate dehydrogenase) mRNA level in human cells (HeLa) by the real-time PCR method with the SYBR Green I detection?

Preferably, the primer pair spans intron, the amplicon size range is 100 – 150 bp and the melting temperature is 60 - 62°C (determined by the Nearest Neighbor Method).

I have tried to design some primers manually or using available public algorithms but all of them displayed 100% similarity to many GAPDH-like human transcripts when BLASTed.

Sorry, if I have overlooked the right thread.

Thanks for any help I can get, Piotr

Maybe you can try these from RTPrimerDB:

http://medgen.ugent.be/rtprimerdb/assay_re...RTPrimerDB_ID=3

but there are so many other GAPDH primer sequences submitted... just have a look.