Hello everyone!
If cell pellet has been stored in -80 for two years. Can RNA be extracted for mi-RNA Seq? Would it be of good quality?
And what's your recommended methods of RNA extraction from cell lines for mi-RNA Seq? Please feel free to comment down below.
Much appreciated!
Using frozen cells for RNA seq can lead to RNA loss due to the rupturing of the plasma membrane during freeze/thaw. You could try single nuclei isolation instead of single cell isolation before sequencing if the miRNA is found also in the nucleus, this would avoid RNA loss since the nuclear membrane is still intact after freezing. A good kit for single nuclei isolation is this one from Invent Biotechnologies: https://inventbiotech.com/products/minute%E2%84%A2-single-nucleus-isolation-kit-for-tissues-cells-20-preps?_pos=1&_sid=c69eff0d3&_ss=r
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