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
- - - - -

Buffer-saturated Phenol

phenol RNA isolation

  • Please log in to reply
1 reply to this topic

#1 Flopilus

Flopilus

    member

  • Active Members
  • Pip
  • 17 posts
1
Neutral

Posted 03 April 2012 - 08:06 AM

How can I prepare a 38% bufer-saturated phenol (v/v) solution? I need it for an Extraction buffer to isolate RNA.

Thanks.

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,799 posts
406
Excellent

Posted 03 April 2012 - 12:33 PM

You can buy it quite easily - many companies sell it, but the most reliable and quickest is Sigma-Aldrich. I would strongly recommend buying it!!!!

Making it is time consuming and relatively dangerous (phenol is toxic, hard to remove, and causes burns). However if you want to proceed with making it, equilibration of phenol is performed by:
1: Remove liquid phenol from freezer and melt at 68 deg C, add hydroxyquinolone to 0.1% (yellow colour gives convenient way to identify organic phase, + is an antioxidant)
2: add equal volume of 0.5 M tris-Cl (ph 4.5) and stir for 15 min. Allow phases to separate and remove upper aqueous layer into appropriate discard (NOT DOWN THE SINK!!!!!)
3: Add equal volume 0.1 M tris-cl, pH 4.5, and stir for 15 min, remove aqueous phase. repeat until pH is 4.5, by measuring the pH of th buffer after each equilibration.
4: when finally equilibrated, add 0.1 volumes of tris-cl, pH 4.5, with 0.2% 2-mercaptoethanol and store in a light proof bottle at 4 deg C for 1 month.

After this time, equilibration will need to be repeated to remove oxidation products.





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

©1999-2013 Protocol Online, All rights reserved.