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??
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#2
aimikins
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Posted 27 April 2009 - 08:03 AM
yeah, sure. that's the easiest way. just make sure the dilution range is nice and wide.
"it is a miracle that curiosity survives formal education" -A.E.
#3
thegene
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Posted 28 April 2009 - 12:06 AM
aimikins, on Apr 27 2009, 09:03 AM, said:
yeah, sure. that's the easiest way. just make sure the dilution range is nice and wide.
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
#4
aimikins
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Posted 29 April 2009 - 01:48 PM
what is your dilution range?
also, can you add a pic of your melting curves?
also, can you add a pic of your melting curves?
"it is a miracle that curiosity survives formal education" -A.E.
