Does incubation with proteinase k before a totalRNA extraction degrage miRNA's?

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Saurabh Mandal

In the case of clinical samples, studies suggest Proteinase K treatment improves RNA quality and yield. It could be miRNA isolation kit that might not be giving the expected result.

you may see once the proteinase K treatment studies of clinical samples.

Article A faster and less costly alternative for RNA extraction of S...

https://www.frontiersin.org/articles/10.3389/fcimb.2021.717068/full

https://www.medrxiv.org/content/10.1101/2020.12.16.20248350v1.full

Article Proteinase K treatment improves RNA recovery from thyroid ce...

Article Detection of circulating miRNAs: Comparative analysis of ext...

Could you please elaborate what is the reason for the higher miRNA yield from your isolation? If I am pointing correctly, then sample id 1--5 and 16-20, which may be 'without proteinase K', and if we compare those samples with others, there is no big difference.

Average estimation describes miRNA comprises ~0.01% of total RNA by weight. Even using standard kit isolation sometime miRNA isolation may not meet expectations. if your samples preparation using a different kit or you have done miRNA-specific treatment and then any difference emerges then that needs further analysis. However, You can analyze the miRNA of interest in these isolated samples using qRTPCR. Then see what happens.