Hello,

I am new to the world of Reverse Transcription PCR and would really appreciate some advice about whether it is the right technique for me or not…

I have the full genome sequence of the bacteria I am working with, I had previously designed and DIG labelled probes specific for the operon I am studying (for northern blotting experiments). BUT…
Is it possible to use RT-PCR to follow expression?
Would the probes need to be redesigned?
(& if so what is the best technique for designing them?)
What company is best to purchase the reagents from?

Sorry if any part of my query is unclear, please do not hesitate to contact me for any further information.
Many Thanks,
Eve

RT-PCR is a great way to follow gene expression profiles. You can use genomic DNA or cDNA. You will need to remake/order primers and probes. I use Taqman from applied biosystems for my master mix and order my primers and probes from biosearch, applied biosystems or biosource. (all 3 companies are good for primer/probes).

Let me know if you have any specific questions. I'd be happy to answer

Hello,
Thanks for your reply. I shall go and look up the products you suggested and formulate some questions!
Cheers,
Eve