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
- - - - -

getting strange data in Luciferase-renilla assay

luciferase-renilla

  • Please log in to reply
2 replies to this topic

#1 gingerx

gingerx

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 21 March 2013 - 05:58 PM

hi guys
I am doing the promega dual luciferase-renilla assay(miRNA testing on UTR)
but my ctrl which for sure was used in other experiments do not go down. Anyway I do not have any desired data about all other samples
renilla reads are really different between different samples but in same cells and I thought it should not differ much between different samples
i got a ratio of Luc per Ren of 1 for my empty vector ctrl but all other samples were higher than 1!
I seeded 70000 cells per well(24well) the next day transfected them and after 24hrs I read the plate
Is there any point that I have missed?any idea?
Thanx

#2 pcrman

pcrman

    Epigenetist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts
67
Excellent

Posted 22 March 2013 - 10:41 PM

i got a ratio of Luc per Ren of 1 for my empty vector ctrl but all other samples were higher than 1!


I am not sure what your specific question is. Of course you may have different L/R ratios in different treatments because treatments may altered luc activity. How about the reading of renilla activity alone? Is it pretty consistent across treatments?

#3 gingerx

gingerx

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 25 March 2013 - 11:17 AM

renilla between replicates show consistency but among different treatments not
u know the upregulation was not the expected result as a similar experiment already has done by sb else in our lab.even my positive ctrl that I am sure about its downregulation does not work




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

©1999-2013 Protocol Online, All rights reserved.