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
- - - - -

Luciferase assay - extreme variation in RLU


  • Please log in to reply
1 reply to this topic

#1 Pkrish

Pkrish

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 01 March 2013 - 08:40 AM

Hi

I am performing luciferase assay with 2 expression vectors+ 1 promoter-luciferase vector+GFP- control vector.

So each time i perform the experiment I get extremely different readings.

Could it be because one of the vectors is not getting transfected, even though i get a pretty decent transfection efficiency. Also, can only the GFP vector be transfected and not the others?

Suggestions and ideas would be immensely helpful

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,733 posts
400
Excellent

Posted 02 March 2013 - 12:01 AM

It is certainly posssible that there are different transfection efficiencies for each plasmid, though if you have optimised this transfection (i.e. amounts of each plasmid and total DNA) then this shouldn't be a problem.

The variations could be due to all sorts of things - you might be better off comparing % change between controls and treatments rather than trying to compare absolute values.




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

©1999-2013 Protocol Online, All rights reserved.