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
- - - - -

Problem with Telomeric Length measurement using RT-PCR


  • Please log in to reply
1 reply to this topic

#1 drchewan

drchewan

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 17 September 2010 - 06:23 AM

Need an advice!

I'm looking at Telomeric length using Telomeric primers developed by cawthorn with some modification by ME Gil and Coetzer, 2004.

I have optimized the assay of Telomere primers and single gene copy 36B4 using Roche Lightcycler 480. I have done validation curve for both the assay and the Regression curve were both more that 0.99. However, the problem lies onto the efficiency of Telomere. For Telomere, I have repeated 3 times with R2 for all of 3 were >0.99, however I got different efficiency such as 2.2, 2.1. and 1.75. For 36B4, The efficiency lies between 1.7-1.9.

I have also tried on the samples, however, everytime I'm repeating the same sample, it give me different value. The value expressed in delta Ct.

I just wonder whether it is possible to get different results due to amplifying repetitive sequence such as Telomere? If not, how this possible? I mean whether it amplyfing primer dimer or unspecific products? can I still use the primers eventhough it give me 2 different value in the same sample?

Thanks,

WA

#2 dtae

dtae

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 59 posts
0
Neutral

Posted 07 October 2010 - 11:52 AM

take a look at:
http://www.protocol-...h__1#entry83844




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

©1999-2013 Protocol Online, All rights reserved.