qPCR - Standard curves (Apr/27/2009 )

Hi mates,

i am trying to do standard curves for a couple of primer pairs that i want to use for measuring the expression levels for a gene (MRNA) after mammalian cells transfection with a plasmid containing the cDNA of the gene of interest.

The standard template with known concentration that i have used for standard curves, is the original plasmid used for transfection with different serial dilutions. Is that correct??

yeah, sure. that's the easiest way. just make sure the dilution range is nice and wide.

aimikins on Apr 27 2009, 09:03 AM said:

Thanks Aimikins,
I have performed two times and in every time i didn't get amplification plots in the qpcr, i feel that, it is not right to use plasmid DNA as a template for qPCR. Do you any idea about that?

Regards

what is your dilution range?

also, can you add a pic of your melting curves?