qPCR amplification - (Mar/24/2014 )
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?
How long is the expected bp length of the product ? Did your gel have an appropriate resolution and running time ?
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.
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?
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?