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
- - - - -

Agarose gel bands smear/ drag problem


  • Please log in to reply
4 replies to this topic

#1 amehac128

amehac128

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 05 November 2010 - 01:58 AM

I am having a problem with my 1.5% agarose gel, where is seems that the bands are dragging???? I have tried fresh TAE buffer for the tank and the gel. I seem to get this problem with my DNA PCR products, of various concentrations and with my freshly made 1kb marker. Any ideas?

#2 BioMiha

BioMiha

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 235 posts
9
Neutral

Posted 05 November 2010 - 02:06 AM

Is it smearing or do you get smiley bands. For the latter I would guess it's the heat. Try running at lower voltage to prevent the gel from heating up. For the former it's usually because of degradation in the sample. Do you have an image of the gel, so we can better see the problem.

#3 amehac128

amehac128

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 05 November 2010 - 03:09 AM

Thank you. I'm not sure how to descibe it really! im running it at 110V for 2.5hrs. I have just run a 2% e-gel with the same samples and not seen this problem run really need to run it longer for the band differences i am looking for. the attached pic has markers in lanes 1 and 20. thanks again.

Attached Thumbnails

  • l.gif


#4 Ameya P

Ameya P

    Rervm Cognoscere Cavsas

  • Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 330 posts
25
Excellent

Posted 05 November 2010 - 11:47 PM

I think the agarose is not polymerizing properly in this case. Give it some more time for polymerization.

NEW!!!!  The Beauty in Symmetry on CoffeeTableScience!!!!

Image copyright: Adrian Koh SF.
Replication of this art is strictly prohibited without express permission of the artist


#5 BioMiha

BioMiha

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 235 posts
9
Neutral

Posted 06 November 2010 - 12:12 PM

I don't think the problem is the agarose. I've had this problem myself. In my experience this happened because voltage was too high or because the salt content of the samples was too high. I am guessing that your loading buffer is too concentrated. Try a different loading buffer.
Just my two cents.
Best of luck.
Miha




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

©1999-2013 Protocol Online, All rights reserved.