Posted 27 April 2009 - 06:50 AM
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??
Posted 27 April 2009 - 08:03 AM
Posted 28 April 2009 - 12:06 AM
yeah, sure. that's the easiest way. just make sure the dilution range is nice and wide.
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?
Posted 29 April 2009 - 01:48 PM
also, can you add a pic of your melting curves?