standard curve for qPCR

[fibonacci]

i'm planning to do a relative quantification and from what i know, i have to perform standard curve for each genes that i used in qPCR. actually, i have about 25 genes of interest. so, do i really need to perform the standard curve for 25 genes??

[CharonY]

Basically yes, as each PCR reaction is different from the other (i.e. different primer/template combo). Sometimes people also just compare the measure value of a sample of interest against a single reference point for a semi-quantitative analysis. However, the danger is that one may be outside the dynamic range.

[jennmitch75]

i'm planning to do a relative quantification and from what i know, i have to perform standard curve for each genes that i used in qPCR. actually, i have about 25 genes of interest. so, do i really need to perform the standard curve for 25 genes??

 

There is a qPCR community at http://qpcr.community.appliedbiosystems.com/index.jspa and it has a white paper that might help answer your questions. You can skip the community and go directly to the whitepaper here http://find.lifetechnologies.com/qpcr-whitepaper/blg

[Dr. Monte Miller]

PCR reactions are notoriously different for each different setup. Different primers, template, mg concentration, temperatures, cycles etc. If all of the parameters are different, and even if most of them are the same, the efficiency of the polymerase and the binding, and the A-T, C_G content will drastically effect the qPCR times.

 

Bottom line yes.

 

mmiller@forensicdnaexperts.com