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
- - - - -

Help with plant DNA contaminants and troubleshooting suggestions

DNA plant DNA contaminants proteins polysaccharides

  • Please log in to reply
No replies to this topic

#1 paula1

paula1

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 04 May 2013 - 10:46 AM

Hello!

I've been having problems extracting DNA from silica gel dried leaves from a plant known to have a serious polysaccharide contamination issue. It also has high quantities of tannins and polyphenols. The resulting DNA is very viscous and the nanodrop profile is catastrophic. I get samples with a 260/280 ratio of 1.3 and a 260/230 of 0.14 (seriously). I use a protocol for obtaining high quality DNA from plants with high quantities of polysaccharides (Russell at al 2010), with sorbitol buffer.

What I want to know is this: do you think I'd get better results by doubling the concentration of sorbitol in my buffer? Also, given the low 260/280, would you advise me to add proteinase K to the buffer as well? The protocol already contains a phase in which the samples are bathed in CIA for 30 minutes, so I think I shouldn't be getting such bad 260/280 ratios. Maybe increasing the CIA bath time instead of adding proteinase K? I don't know, I'm confused. Any ideas on why this is happening and what could solve my polysaccharide + protein problem?

Thanks a lot! =)


PS: if it helps, I can add the nanodrop results for more information.





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

©1999-2013 Protocol Online, All rights reserved.