I suggest dissolving your RNA in Tris-EDTA and add 1 v chloroform before centrifugation. This will remove phenol, but you will lose about 20% RNA yield too.

NOTE: If your samples have a large amount of phenol you may see absorbance at about 265-270 nm.

Thanks a lot for your answer.

But the reported yield in my samples are low ,so It s risky to lose 20% of them.

As Mehmet wrote, use chloroform although you will lose some RNA. The phenol presence in your sample is a major problem for further downstream applications.  If you don't want to use chloroform, you can precipitate your RNA overnight at -80 and wash afterwards with 75 % alcohol but as well there will be some lost during manipulations but better lose some RNA than having phenol contamination. Good luck!

You will never remove low amounts of phenol with chloroform. The only way is the classic precipitation with acetateNa/ethanol at -20•C overnight. It is important to pellet in a cold centrifuge (-4 is enough) and then washing pellet twice with cold ethanol. Centrifugation at cold temperature leaves the pellet sticky to the tube wall. It is important to remove completely the alcohol in a liophylizer before TE addition to solubilize the pellet. You can easily find detailed protocols in Maniatis  book or   of Molecular Biology Current Protocols.

Hello Yordan Muhovski Mehmet Karaca I am in a similar situation. I’d like to know if I use 1v of chloroform (eg. 500ul of chloroform and 500ul of diluted RNA sample), what should be the volume of isopropanol I should use in the downstream step (i.e. when mixing the aqueous layer with the isopropanol)? Thank you!!