I am detecting ~15 different miRNAs in serum samples. For normalization it seems most people use spike-in (cel-miR-39) or endogenous small RNA.
I see various studies normalize to multiple endogenous or spike-in miRNAs using geotmetric mean or other methods, but I don't see any studies that actually normalize to both spike-in or endogenous small RNA.
I want to use both spike-in and endogenous small RNA for normalization. Can this be done? Please refer me to some literature where people have done this.
Thanks,
SC
Yes, you can use a spike-in and a endogenous reference to normalize your samples. The qbasePLUS algorithm enabled normalization to more than one reference gene and also employed inter-run calibrator signals to minimize inter-run variability distinguishing it from the ΔΔCt method. All data were expressed as calibrated normalized relative quantities (CNRQs)
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