gene-specific RT-PCR - (May/29/2009 )

Hello Everybody,

I want to design a gene-specific primer for RT-PCR. What do I need to keep in mind?

Thanks a lot
Hairpin

-Hairpin-

Do you mean doing RT using gene specific primer? I think all RT-PCRs are gene specific.

-pcrman-

pcrman on May 30 2009, 12:03 AM said:

Do you mean doing RT using gene specific primer? I think all RT-PCRs are gene specific.

I do not mean real time PCR. I want a primer for "reverse transcription PCR. Normally you use oligo dTs or random primers. But the mRNA I am looking for is very seldom. I thought that I can increase the efficiency for that gene by using a gene specific primer.

-Hairpin-

I don't think that pcrman thought you meant real time........... and the RT-PCR reactions I have performed have all been gene specific, it is not an uncommon thing (I would actually suspect that the vast majority of RT-PCR reactions are using gene specific primers rather than the method you were describing....but I could be wrong, it has been known to happen )

I have a question for you though, how were you hoping to find your mRNA if you weren't going to use gene specific primers?? How would you know it was the mRNA you were looking for?? I am confused......

If you give us a bit more info about what exactly you will be doing, and any concerns you have, we might be able to help you better.

-leelee-

If you give us a bit more info about what exactly you will be doing, and any concerns you have, we might be able to help you better.