Dear everyone,
we are planning to perform long-read RNAseq on RNA extracted from human post mortem material (frontal cortex). as could be expected, the RIN is quite variable (range: 2,3-7,7). we will probably have to compromise between quality and number of cases we can include in the study. which is the minimal RIN to generate meaningful data from long-read RNAseq?
many thanks in advance,
Chiara Valori
RIN above 7 was fine for my research but mine was extracted using bone marrow aspirates. Anyways if your sample is precious and you have the budget, you can try with lower RIN number and perform your internal validation whether you can proceed with RNA sequencing.
In my experience, the suggested protocol for your intended kit states such requirements. In addition, I would suggest checking the lengths of your DNA by bioanalyzer or something similar. You should have a significant peak at long lengths and low peaks at short lengths.
It is not possible to give you a definitive answer. It depends on what you target with your long-read experiment as well as on the kit/technology you want to use. You might have to do a few extra steps to get rid of very small fragments to at least attempt to get some long molecules sequenced.
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