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
- - - - -

real time qRT-PCR and low expression gene

low expression gene reat time qRT-PCR

  • Please log in to reply
2 replies to this topic

#1 George250

George250

    member

  • Active Members
  • Pip
  • 15 posts
0
Neutral

Posted 11 September 2011 - 11:39 PM

Hello,

Your advice would be much appreciated. One of the genes I am working with (imprinted gene) is expressed at extremely low levels, so reat time qRT-PCR fails to detect any transcripts. Can anyone suggest any other ways to detect few transcripts (other than performing pre-amplifaction)?? Is there any other PCR method (nested, conventional etc) that is more reliable than real time qRT-PCR?

Thank you very much

#2 vitalgene

vitalgene

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 53 posts
0
Neutral

Posted 12 September 2011 - 07:12 AM

at what cycle number do you see the expression (start) of this gene ? and upto what amt of starting cDNA (in ng/reaction) have you tried to conclude that it fails to express in standard condition?

#3 George250

George250

    member

  • Active Members
  • Pip
  • 15 posts
0
Neutral

Posted 12 September 2011 - 02:36 PM

thanx for your reply, i see expression after clycle 36.All my samples have standard number of cells (always less than 1800 cells). When I use around 1800-2000 cells I have no expression. If I do the same and use RNA that was purified from >6000-8000 cells I get some amplification but the expression is very very low. But the point is to do the analysis on small amounts of cells




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

©1999-2013 Protocol Online, All rights reserved.