I am trying to to interpret miRNA expression in certain sample. I have only raw ct value. It would be very nice to include concentration in relation to ct value.. has anyone done it??? help and suggestions will be highly appreciated. thank you
Including a standard curve of your miRNA of interest--different concentrations of a synthetic oligo--allows accurate copy number estimation.
did you mean the absolute quantification??
If you have time please check the paper
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0084686
where in Figure 4A they used ct and miRNA concentration.
@ Kenneth
In this paper, the "miRNA concentration" is provided as "arbitrary units." Although the materials and methods section is not extensive, this calculation is relative and is presumably based on an ideal (100% efficient) reaction and dCt values. A rough estimate, then; conclusions about the actual concentration would require a standard curve and/or, potentially, a digital approach.
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