Hi All,
I'm trying to extract RNA from plasma in order to isolate miRNAs by using Qiagen's miRNeasy. However, I'm getting RIN values of 1 and 2 on an Agilent Bioanalyser.
Are RIN values neccessary when working with miRNA? And should I use exosomal RNA instead? Thanks
Hi dear, The calculation of the RIN is based on the ribosomal RNA and has nothing to do with miRNA. If you have a lox RIN it means that your ribosomal RNA and the mRNA are degraded but as the miRNA are very small molecules they may be OK in your preparation.
For me RIN is very importan, as you know miRNAs are small and if your RNA sample is degraded you can isolate degradation products together with miRNAs and carry problems with cloning and sequencing. Try to get a very high quality total RNA.
Dear Nohemi,
For cloning and analyzing miRNA it is not a problem to have a degrade rRNA and mRNA because you have to amplify your miRNA using specific primers. extracting miRNA there are different kist that help to eliminate the other types of RNA even if the RIN is around 10.
Plasma samples have very low RNA concentrations and when you have samples with very low concentration assessing quality with OD or Bioanalyser might not be suitable.
Exiqon have some nice guidelines for working with biofluids and microRNA, and you can also get some tips on sample handling (like not to use heparin when isolating micrRNA) etc:
http://www.exiqon.com/ls/documents/scientific/microrna-serum-plasma-guidelines.pdf
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