Hello everyone,
I'm new to Real-Time PCR and I'd like your insight on this. I have generated standard curve by doing 1:10 serial dilution in triplicate. However, the result obtained, of which R value is to determine the efficiency of the assay is poor (R> 114%) and there are only three points out of 10 that fall on the line. Should I rerun test on the rest of the deviated points or repeat the whole procedure again?
Many thanks
You'll have to repeat all of it.
But it's normal to have limits of your useful detection range. Too few copies and you don't get efficient amplification. Same can happen for too many copies of the template.
The R-square value tells you have precise & accurate repeatability for technical replicates.
The slope of the Ct vs template concentration tells you the reaction efficiency.
Both of these measures are important.
Good luck!
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