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
- - - - -

incorrect/smaller band size for real time PCR


  • Please log in to reply
1 reply to this topic

#1 labpeace

labpeace

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 06 November 2010 - 01:02 AM

Are real time PCR products shorter in size as compared to standard PCR ? I am expecting a band size of 248 bp but I am getting 100bp on a gel. My NTC is negative and there are no primer dimers. What is the cause for getting a smaller band size ? Any advice would be appreciated.

#2 ivanbio

ivanbio

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 116 posts
7
Neutral

Posted 06 November 2010 - 12:24 PM

Real time PCR products are not smaller than regular PCR products. Real time PCR assays are typically designed as smaller amplicons, but if you designed it to be a 248 bp amplicon, then it should run around that size on a gel.

Of course it all depends on what you are comparing it against. If you are running a size ladder that uses a different loading buffer than the one you use to load your real time PCR product, and you did not clean your real time PCR product (i.e. your real time PCR reaction still contains SYBR Green), then your PCR product will likely run much different in size on a gel.

Ivan
Carlsbad, CA




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

©1999-2013 Protocol Online, All rights reserved.