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qPCR amplification - (Mar/24/2014 )

hi.

I'm wondering, is it possible to have nice amplification plot in qPCR but no band observable when the pcr product is run on etbr stained gel?

-Angeline-

How long is the expected bp length of the product ? Did your gel have an appropriate resolution and running time ?

-Tabaluga-

It's very possible to have nice amplification plot (usually high Cts) of primer-dimers.

These are really small and can just run out on ordinary gel, higher percentage gels are needed and mostly the dimers are visible as a thick smear.

-Trof-

I understand that there is a possibility of primer dimers if the ct is high.

But, is it possible to not get band on etbr stained 1.5% agarose gel if the ct value falls within the range of 8 to 15? 

-Angeline-

Are you sure that your gel works? (or if you don't repeat it, that it worked at that moment?)

Can you run other "working" real-time PCR side by side and put both also on gel?

 

Did you do a melting curve analysis of that product?

-Trof-