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
- - - - -

phosphorylation of primers using PNK


  • Please log in to reply
1 reply to this topic

#1 marikab

marikab

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 22 June 2010 - 12:20 AM

I phosphorylated my primers using T4 polynucleotide kinase - don't think it worked... Read somewhere that during the synthesis/desalting of primers they use ammonium hydroxide. Ammonium ions inhibit PNK. Any suggestions for removal of ammonium ions from primers?

#2 tfitzwater

tfitzwater

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 96 posts
3
Neutral

Posted 22 June 2010 - 06:44 AM

1. The ammonium hydroxide is evaporated off during drying.
2. It takes a high concentration of ammonium to inhibit PNK. (Yes, I have performed the titration.)
You are unlikely to encounter the concentration levels necessary to inhibit the enzyme.




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

©1999-2013 Protocol Online, All rights reserved.