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
- - - - -

PCR comparison of gene expression


  • Please log in to reply
4 replies to this topic

#1 vetticus3

vetticus3

    Princess

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 144 posts
9
Neutral

Posted 13 December 2004 - 04:24 PM

Hello,
can someone shed some light on this topic for me... if someone were to set up a PCR based experiment to compare the expression of two different genes.... this would be a real time PCR? and it would require primers specific for those two genes, and another set of primers for a control (ie GAPDH etc)?
is there another way to compare the expression of two genes?

thanks

vetticus

#2 green

green

    member

  • Active Members
  • Pip
  • 24 posts
0
Neutral

Posted 13 December 2004 - 05:05 PM

You are right, vetticus.

For expression analysis, any technique that involves amplification is not reliable. Hybridization analysis such as Northern and Western are much better than PCR based methods.

#3 vetticus3

vetticus3

    Princess

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 144 posts
9
Neutral

Posted 13 December 2004 - 08:53 PM

cool, THANKYOU!! green,

what's used in a clinical setting? do pathologists use RT-PCR or nothern analysis? if you've been diagnosed with cancer, and it's been cut out, and a sample is sent of to pathology for it's characterisation... and a part of it is to do an analysis of gene expression..... would they do the quicker, though questionable RT-PCR.... or a time consuming northern? or are they moving on towards microarrays?

#4 green

green

    member

  • Active Members
  • Pip
  • 24 posts
0
Neutral

Posted 13 December 2004 - 09:29 PM

That would be a different story if you are talking about clinical setting. Immunohistochemistry or tissue microarray (if there are many samples) would be the way to go. I don't think cDNA microarray at current stage will yield any meaningful data for clinicians to make decisions.

#5 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,522 posts
371
Excellent

Posted 14 December 2004 - 07:47 PM

Hi

Real time PCR is a validated method for expression analysis of genes, though there are a number of caveats, such as the choice of a control gene, usually people use a gene that has consistent expression throughout the body or tissue of interest such as GAPDH or Beta-actin, though some advocate the use of 18s RNA.

In fact real time PCR has now largely replaced northern or western blotting for expression analysis due to the greater sensitivity of the PCR based system, and papers these days often expect to see real time results rather than the other methods.

Check out this web site for more info: http://dorakmt.tripo...s/realtime.html




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

©1999-2013 Protocol Online, All rights reserved.