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
- - - - -

Selective removal of standards post qPCR Run

qPCR standards

  • Please log in to reply
1 reply to this topic

#1 schookp

schookp

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 05 June 2013 - 08:33 AM

Hello

I was curious as to the rules for removing values from a standard reaction in a qPCR run. In qPCR runs where the effciency is just outside the 90-110% range and removing one standard dilution was that non linear relative to the other standards or the removal of one replicate within standard brings the run back within the 90-100% range, what is the rule for doing so? In this case the test being done is absolute quantification. Obiviously the data output will be altered when standards are adjusted, but unless subtle changes are being examined does this really matter?

Thank you very much for any information that can be provided. I want to make sure that any steps taken are within the norms of qPCR experimentation, and I feel that this is a good place to get that feedback.

#2 jerryshelly1

jerryshelly1

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 309 posts
29
Excellent

Posted 05 June 2013 - 08:56 AM

I do this. I always change the threshold manually, kicking out any outliers. Just make sure you adjust your control thresholds to match your experimental samples.





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

©1999-2013 Protocol Online, All rights reserved.