You know, there are many legends about RNA extraction, including not breathing nearby :) I'll tell you that I have isolated RNA in dirtiest and dustiest places and I have never had a degraded RNA, except when the tissue itself was compromised (like post-mortem tissue). So, don't waste your time on DEPC, just optimize your extraction procedure.

This will help you

http://genomics.no/oslo/uploads/PDFs/workingwithrna.pdf

If there is somewhat difficult in using of DEPC, You just prefer autoclave for sterilization.

You know, there are many legends about RNA extraction, including not breathing nearby :) I'll tell you that I have isolated RNA in dirtiest and dustiest places and I have never had a degraded RNA, except when the tissue itself was compromised (like post-mortem tissue). So, don't waste your time on DEPC, just optimize your extraction procedure.

Thank you Vijay and Leonid.

It is not necessary to treat your tips and tubes with DEPC, as long as sterilization at about 200 ℃.

Add 0.1% DEPC in ddH2O. Mix it with magnetic strirer for about an hour in dark. Dip all the tips and tubes completely merge in 0.1% DEPC water overnight in dark. Dry them and then autoclave. All condition must be sterile such as use proper gloves and clean the surface with 0.01% autoclaved DEPC H2O.

The link suggested by Vijay Gahlaut also helps.

All the best!

Thank you very much Xiaoyi and Saurabh.

You must use new tips and tubes for any Nucleic acid isolation. Fill them up in empty tip boxes with gloved hands. Wrap them properly and autoclave them.

Shefali

I agree with Shefali Madam. Instead of going through the laborious task of treating the tips and tubes with DEPC treated water, drying etc., I suggest you to use certified DNase, RNase, Pyrogen free tipes, which are available with all the major vendors now a days and will not cost too much. And please note that you need not autoclave these nuclease free plasticwares. Just contact your plasticware vendor.

Best wishes,

SD

@ Sefali Desai, yes we are using all new tips and plastic wares. I was little bit skeptic about moisture retention by the filters in the filter tips. Thank you for your suggestion.

@ Sibnarayan Dutta, we use certified DNase, RNase, Pyrogen free tips. About autoclaving of plastic wares, we will take note of that. Thank you for answering

As everyone else has said, RNA extraction does not need to be done as carefully as most people seem to think. Simply autoclave your tips and that is enough.

DEPC is carcinogenic, I recommened not to use it. By RNase free plasticware.