Hi I am using RNAse Inhibitors (applied biosystems), which are enzyme-based and are meant to add during cDNA synthesis phase as per its manufacturer policy (1ul to each 20 ul RT reaction).
But, can I also add it just after the RNA extraction into my RNA elutes (FFPE-based) in the same ratio and store that RNA at 4 C for some time (like 24 hours) before carry on the reverse transcription phase? Just to avoid any further RNA degradation of FFPE extractions...Any informative thoughts!!!!!
I would say that you can add RNase inhibitors to your sample, however, keep in mind that if there is already partially degraded RNA (the quality/integrity of RNA isolated from FFPE is always lower) it will not save your samples. Most importantly, I would not leave RNA at 4 C, not even only O/N. Instead, put it to -20C; that way you can keep it 1-2 days (I do it usually this way and never had a problem).
Good luck!
- I would say you probably can but you shouldn't need to if your RNA it totally pure and therefore devoid of RNAses
- In terms of possible downstream contamination when you perform an enzymatic reaction, e.g. RT then yes I would add to that reaction
- In fact it might be btter to store your RNA minus things like RNasin and only add to aliquots that you are performing down stream reactions with e.g. cDNA synthesis
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