Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > PCR - Polymerase Chain Reaction Forum > Real-Time PCR and Quantitative PCR Forum
Register Search Today's Posts Mark Forums Read

Real-Time PCR and Quantitative PCR Forum Real-Time PCR and Quantitative PCR Forum


TaqMan Gene Expression Assay Bewilderment

TaqMan Gene Expression Assay Bewilderment - Real-Time PCR and Quantitative PCR Forum

TaqMan Gene Expression Assay Bewilderment - Real-Time PCR and Quantitative PCR Forum


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 02-22-2011, 09:52 AM
Pipette Filler
Points: 75, Level: 1 Points: 75, Level: 1 Points: 75, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2011
Location: Warwick, UK
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default TaqMan Gene Expression Assay Bewilderment



Hi,

This is hopefully a really basic error I'm making, but I am very new to this.
I'm using the TaqMan Gene Expression Assay to measure thrombomodulin expression in endothelial cell lysates. I'm using 18s as the endogenous control. My results are all over the place. Even untreated samples return hugely different dCt values (4 cycles apart). The strange thing is that the curves for the thrombomodulin probe are always pretty much superimposed. All the variation comes from the 18s curves which are widely spread (4 cycles apart!)

I can accept that the 18s curves will separate a little due to pipetting error, but it seems a mighty coincidence that every time the thrombomodulin curves are closely packed.

I hope this makes sense to someone!
Reply With Quote
Reply

Tags
assay , bewilderment , expression , gene , gene expression , taqman


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Human Cytome Project - Update 24 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 1 08-01-2010 02:18 PM
New Saccharomyces Sequences 11/27/04 Mike Cherry Yeast Forum 0 11-29-2004 12:39 AM
New Saccharomyces Sequences 09/08/04 SGD Sequences Yeast Forum 0 09-13-2004 10:07 PM
New Saccharomyces Sequences 08/11/04 SGD Sequences Yeast Forum 0 08-12-2004 12:26 AM
New Saccharomyces Sequences 05/19/04 SGD Sequences Yeast Forum 0 05-23-2004 04:06 PM


All times are GMT. The time now is 03:56 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.16155 seconds with 16 queries