qPCR question (no results?) - (May/31/2013 )
Hi I'm having trouble with qPCR's. I just finish my first practice qPCR with just one sample (an RNA control from a kit my mentor no longer needed). After I ran my qPCR in the machine however, there was no amplification curve at all. The data screen was totally blank! What could I be doing wrong? I thought maybe it's because the concentration of the RNA was too low? It's only 1ng/ul. My mentor told me to dilute the actual RNA samples to 25ng/ul once I move on from practice qPCR's because their concentrations are all too high (>100ng/ul)
I'm using taqman assays, if that makes a difference.
Take your PCR and run it on a gel - this will tell you if you have a product or not.
Have you done normal PCR before - if so do those work? If not - practice on those first (it's much much cheaper), then you shouldn't have any problems.