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
- - - - -

3' Race PCR


  • Please log in to reply
No replies to this topic

#1 nirupakl

nirupakl

    member

  • Active Members
  • Pip
  • 11 posts
0
Neutral

Posted 24 February 2010 - 01:33 AM

Hi,

I have gel eluted my 3'race product.Upon subsequent dilution of the gel eluted product and reamplification using nested primers (nested gene specific primer and nested universal primer supplied by the company), I have been getting numerous bands for 3'race pcr alongwith my band of interest.The sample run on a agarose gel looks like a ladder of DNA. I also tried using nested primers internal to the sequence and i get a clean single band apart from the primer dimer.
I have tried playing with temperatures, tried Touch down pcr and got fresh stocks of primers to sove contamination issues if any. The problem seems to persist.
Can i have any suggestions please?




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

©1999-2013 Protocol Online, All rights reserved.