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qPCR question (no results?)

PCR qPCR real-time missing data amplification curve

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#1 LQJ

LQJ

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Posted 31 May 2013 - 10:37 AM

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.
Thanks!!

#2 bob1

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Posted 01 June 2013 - 01:28 AM

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.





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