I am extracting RNA from mouse tissue and would like to send the samples for sequencing, particularly looking for miRNAs in the samples. How useful and resourceful is the RIN value when the samples are measured with Bioanalyzer? Is there also a threshold for RIN value before sending for sequencing?
Hi Ketharini
RIN is an indication of the integrity of RNA. It is important for quality control of every sample of total RNA (where you can localize the ribosomal RNAs). Of course that a bad RIN in a sample, will indicate degradation, and presence of small RNA fragments that will be subsequently purified and sequenced in the small RNA-seq experiment.
So, the better the RIN, the more guarantee you have to see only small RNAs in your sequencing and not degradation products.
Best
Hi Ketharini,
I would add some more suggestions. First, to limit the activity of RNases, it would be recommended to stabilize the samples (e.g., using RNAlater solution) - it could standardize the sampling, and limits fears of bad RINs (this number has exactly nothing to say about miRNAs, but indirectly adds some view of the samples quality). Note also that miRNAs may be less subjected to degradation than other RNAs, but this may vary.
If concerns persist, testing of selected miRNAs performance using qRT-PCR may be applied. The lower Cts, the better quality.
Good luck,
Luděk
depending on the protocol used for library prep, the people working in the lab, and the nature of the samples used (i.e Urine, blood is very different from tissue in terms of RIN), RIN should be high as possible but I have personally gotten excellent results (80% miRNA reads) from RIN 4 samples
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