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
- - - - -

dna degradation


  • Please log in to reply
4 replies to this topic

#1 avichai

avichai

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 24 May 2009 - 05:08 AM

hello,
I'm working on Lisianthus (plant from the family Gentianaceae).
recently I have a problem of a genomic dna degradation, the DNA was produced using chloroform-octanol it looked O.K at the begging but after a few weeks in -4 it was a complete mess.
any suggestions why it could happen?
what can I do to avoid it?

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,733 posts
400
Excellent

Posted 24 May 2009 - 04:17 PM

Did you resuspend your DNA in 10 mM tris pH 7.0-7.5 or TE buffer? If you used just water, the DNA will undergo acid hydrolysis, degrading it. Otherwise it could be that you have DNase contamination of your samples (maybe in the tubes).

#3 avichai

avichai

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 24 May 2009 - 11:45 PM

Did you resuspend your DNA in 10 mM tris pH 7.0-7.5 or TE buffer? If you used just water, the DNA will undergo acid hydrolysis, degrading it. Otherwise it could be that you have DNase contamination of your samples (maybe in the tubes).

the DNA was resuspended in TE buffer and the tubes were autoclaved...

#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,733 posts
400
Excellent

Posted 25 May 2009 - 04:37 PM

Unusual, perhaps there are some DNases coming through your extraction proceedure. It could be that there is some DNase in one of your reagents otherwise - commonly in the water used to prepare solutions.

#5 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,475 posts
251
Excellent

Posted 26 May 2009 - 04:36 AM

Most plants have phenol compounds which are difficult to remove with standard preps. I'd recommend a CTAB prep rather than only a phenol/chloroform prep for plant tissue.




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

©1999-2013 Protocol Online, All rights reserved.