Dear all, I'm looking for a proper way to analyze my miRNA qpcr data. The study was aim to screen 10+ miRNAs which have been reported to have strong relationship with a certain disease in the clinical cohort. I have to admit that it is indeed a poorly conceived study with only one reference gene included and without PCR amplification determination. The genorm stability shows that this reference gene is at the least stable side. Should I stick to normalize to this unstable reference gene although it may cover the true biological significance? Or can I treat all the genes as candidate reference genes and choose the optimal reference genes to be normalized by performing the genorm stability ranking? Thank you!
Hello Minxia ,
one way to normalize microRNA data when you don't have a proper normalizator ( as it seems in your case), it's to use the global mean. It means that you can normalize your mir ct sample on the mean of all the microRNA you run for that sample. Another way is to select and run other normalizators.
Hope it is usefull,
Goodluck
Claudia
Hi,
I agree with Claudia, but be aware that if all of your miRs are down-regulated (or up-related) using a global mean may mask any real changes you might have.
This approach is more often used on array-type data where you would expect the overall average to be stable.
Hi,
If most of your miRNAs are up- or down-regulated you can always calculate the absolute concentration of your miRNA using a standard curve.
Good luck!
Alfonso
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