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
- - - - -

PCR Primer Design with restriction sites


  • Please log in to reply
1 reply to this topic

#1 rajgene

rajgene

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 34 posts
1
Neutral

Posted 21 December 2004 - 12:35 PM

HI GUYS;
i designed a primer for a promotor region of fcerg1 gene to study its snps effects via reporter gene study.
i did a epcr of the primers from ncbi and it matched perfectly to the corresponding locus, but after i added a restriction sites to the forward and reverse primer mul I and Xho I respecitve÷y for PCR cloning in my PGL2 basic vector, the primers dont score a epcr hit.
is this a serious problem.... will it affect the actual amplication of my product during real PCR.
please explain me how to proceed?
thxx
raj

#2 pcrman

pcrman

    Epigenetist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts
68
Excellent

Posted 02 January 2005 - 02:40 PM

No, it won't. Because you have added some bases to the 5' end of both primers, the program can no longer find a match for the new sequences. So just go ahead with your PCR.




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

©1999-2013 Protocol Online, All rights reserved.