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
- - - - -

how to get rid of non specific bands in PCR


  • Please log in to reply
1 reply to this topic

#1 de ifz

de ifz

    member

  • Active Members
  • Pip
  • 13 posts
0
Neutral

Posted 29 June 2011 - 07:36 PM

Hello guies!! I am quite new to this work. m optimizing a nested PCR protocol in which i am encountering few nonspecific bands along with the required PCR product. Please help me to know where the problem lies?? Is my Mg++ conc. is high?

#2 Ameya P

Ameya P

    Rervm Cognoscere Cavsas

  • Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 330 posts
25
Excellent

Posted 29 June 2011 - 10:03 PM

Please provide a picture for your problem. It makes it easier to make suggestions that should work.

Reducing Mg++, increasing annealing temp, increasing/ decreasing extension time and temperature are different suggestions that can be made depending on where your un specific bands are.

For bands lower than your band of interest, increase annealing temp, for those higher, decrease extension time.

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





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

©1999-2013 Protocol Online, All rights reserved.