Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Does unautoclaved DEPC inhibit northern blotting?


  • Please log in to reply
2 replies to this topic

#1 yekaterina poloz

yekaterina poloz

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 23 August 2010 - 12:55 PM

I use DEPC treated H2O for all of the buffers from electrophoresis to immunodetection during northern blotting. If I stop autoclabing DEPC treated H2O or buffers made from it, will it have a negative impact on the blots? I know carboxymethylation of purines by DEPC inhibits translation but what about migration in the gel, RNA:RNA or DNA:RNA hybridization or immunodetection? It's not that I am lazy but we've all of the sudden have a very limited access to an autoclave.

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,784 posts
405
Excellent

Posted 23 August 2010 - 02:50 PM

DEPC will break down in the presence of water and heat, you can just heat the solution for a while (I don't know how long would be suitable). The major issue is of course that DEPC is quite toxic, so I am not sure that you would want it being realeased into the lab space every time you open the bottles for you to breathe in.

#3 btc8

btc8

    member

  • Active Members
  • Pip
  • 11 posts
0
Neutral

Posted 26 August 2010 - 06:43 PM

I use DEPC treated H2O for all of the buffers from electrophoresis to immunodetection during northern blotting. If I stop autoclabing DEPC treated H2O or buffers made from it, will it have a negative impact on the blots? I know carboxymethylation of purines by DEPC inhibits translation but what about migration in the gel, RNA:RNA or DNA:RNA hybridization or immunodetection? It's not that I am lazy but we've all of the sudden have a very limited access to an autoclave.


You need to autoclave it to inactivate the trace DEPC. DEPC will react with anything that has a -N, -O, or -S, so it may screw up your RNA bases. I would autoclave to be safe, especially if you're using it to store RNA in.

-b




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.