Hello!
I am looking for reliable endogeneous controls for mirna from human and rat plasma. As I understand, use of small nucleolar miRNAs as housekeeping RNAs should be avoided. And the adding of c.el. spike-in controls only uniformity of RNA isolation and reverse transcription. What is the best strategy to normalise miRNA plasma level?
Hello,
Naive answer since i'm not in the field.
If you can assume the individuals have the same circulating volume and you take the same volume for the sample to test, I'd keep it straightforward and normalize the abundance of the detected miRNA to the plasma volume.
If you need to express it more accurately than say arbitrary units per µl plasma you can consider absolute quantification.
Or maybe it is accepted to extract the total RNA from an aliquot of whole blood before taking the plasma and normalize to a house keeping gene of the circulating cells as long as you can assume their nb is the same across individuals (i.e no one has an ongoing inflammation)
Best,
Daniel
Hi,
I suggest to read the article below, it describes how to control the expression of miRNA.
Article Post-transcriptional control of miRNA biogenesis
Hope this may help you.
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- Biological Science
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- Nucleic Acids
- RNA
- Small RNA
- microRNA