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
- - - - -

Genomic DNA quantification using the NanoDrop


  • Please log in to reply
3 replies to this topic

#1 seanspotatobusiness

seanspotatobusiness

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 175 posts
7
Neutral

Posted 25 September 2014 - 08:43 AM

I read that genomic DNA quantification using the NanoDrop may be inaccurate due to the lengths of DNA strands but I used a genomic DNA purification kit which renders the DNA into fragments no larger than 100 KB. Is this too long for accurate NanoDrop readings?



#2 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,505 posts
253
Excellent

Posted 25 September 2014 - 01:52 PM

There should be no problem with fragments that size.



#3 Bio-Lad

Bio-Lad

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 85 posts
16
Good

Posted 26 September 2014 - 04:49 AM

Out of curiosity, at what size does this become an issue? I mostly use our nanodrop for quantification so it would be good for me to know :-)

Try pLOT, the a free plasmid mapping tool!

http://plasmidplotter.blogspot.com/


#4 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,505 posts
253
Excellent

Posted 26 September 2014 - 06:58 AM

With DNA that is of the megabase or greater size, the strands physically clump together, which is why you can spool them with a glass rod. Although they are dissolved, the solution is not uniform in concentration, so pipetting it is problematic, especially for small volumes. Shearing the DNA, or using columns shortens the strands and makes the solution more  uniform in density. I'd say 100 kb is about the point at which this is a major factor, but if you told me it was several fold larger or smaller, I would not be shocked.






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

©1999-2013 Protocol Online, All rights reserved.