What's the maximum temperature at which small RNA (100-200nt) will lose its secondary structure without complete degradation?

22 March 2019 1 9K Report

I have some RNA transcripts about 130nt long on a Bioanalyzer run, and they should be closer to 100nt. I think it's due to secondary structure. What's the maximum temperature at which I can incubate the RNA in nuclease-free water to remove secondary structure without degrading the backbone?

Hasan Issa

RNAs are very stable at high temperatures in the absence of nucleases. You can safely denature at 95°C. I heated RNAs up to 99°C for 30 minutes and they were absolutely fine. However, I would not use nuclease-free water, instead a buffer containing EDTA would be better.

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