RNA isolation is not that complicated, nevertheless the purity and the quantity depends on your pipetting skills. I have been using TRIzol reagent (also called RNAiso) for RNA isolation. It separates RNA, DNA and protein into three layers via a liquid-liquid extraction principle. Due to acidic nature of the reagent, RNA will remain in the top aqueous layer of the mixture (Trizol+sample) after centrifugation (several steps are there, see link). You can now carefully pipette out the aqueous layer to isolate the RNA. The aqueous layer is followed by an 'organic phase' (thick 'milky' phase), So, while pipetting out the aqueous layer, make sure you are not taking some (or substantial) amount of 'organic phase'. As, I said before, the purity depends on your pipetting technique.

This is link to a guide that will help you:

https://tools.thermofisher.com/content/sfs/manuals/trizol_reagent.pdf

But, If you are looking for a conventional way of RNA isolation the results may not be reproducible, could be highly erroneous and almost always the purity is not up-to the mark.

Hope, this helps.

Hi Dilshad,

While using RNA extraction using Trizol reagent, take the clear upper fraction from the Trizol extract and put it down a QIAGEN RNeasy column. Wash with the kit buffers and elute with 30-50ul RNase free water depending on how concentrated you want your RNA to be. Thus, you can get good 260/280 ratios pretty quickly. Best wishes.

intersting question