Hi everyone! do you have any piece of advice about RNA extraction from microvescicles? I usually recover MVs from human monocytes supernatant. I tried different protocols, like trizol and columns but I've never managed to detect RNA through bioanalyzer...thankyou!
Have you tried to perform RT and run a PCR on your extracts? Even though you are below the detection limit of the bioanalyser you may still be able to produce amplifiable cDNA if there is any RNA there at all.
We were extracting cell-free RNA from plasma samples of patients. Although the concentrations were not very high, we were still able to detect miRNA molecules with qPCR.
We used 50 ug/ml glycogen as a co-precipitant and then extracted total RNA with trizol. Then we purified the RNA with columns. The glycogen did help us decrease Ct values by 1-2 in qPCR.
You can try RNA isolation kit of thermo fischer,link is mentioned below.If you do not able to detect through bioanalyzer, still you can perform RT and run a PCR, as per Nicholas Robert Howell said. I did the same and got the desired result.
https://www.thermofisher.com/order/catalog/product/4478545
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