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
- - - - -

proteinase treatment


  • Please log in to reply
2 replies to this topic

#1 lyok

lyok

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 206 posts
6
Neutral

Posted 22 June 2013 - 11:11 AM

Dear all,

I am trying to collect DNA from samples that require a pretty long proteinase K treatment.
I was wondering if I could do this "incubation" overnight (+ 14 hours of incubation). Or would this perhaps damage the DNA?

The incubation happends at 57°C , so I wonder if this is bad? (since I always hear you need to keep DNA at low temperatures to keep it stable..)

thanks in advance

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,831 posts
415
Excellent

Posted 22 June 2013 - 01:55 PM

DNA is a pretty stable molecule, the major damage would result from DNases, which will be largely inactive at 57 and the proteinase K and salt conditions in the digest should also help inactivate the DNases. It is common practice to do DNA extractions from mouse tail tips overnight under conditions pretty similar to those you describe.

#3 lyok

lyok

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 206 posts
6
Neutral

Posted 22 June 2013 - 11:19 PM

DNA is a pretty stable molecule, the major damage would result from DNases, which will be largely inactive at 57 and the proteinase K and salt conditions in the digest should also help inactivate the DNases. It is common practice to do DNA extractions from mouse tail tips overnight under conditions pretty similar to those you describe.


Ok, good to know.
The reason I asked is because I did read that sometimes they advice not to let it stand for more than 8-10 hours.




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

©1999-2013 Protocol Online, All rights reserved.