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


Primer Dimer problems in real-time PCR

Primer Dimer problems in real-time PCR - Real-Time PCR and Quantitative PCR Forum

Primer Dimer problems in real-time PCR - Real-Time PCR and Quantitative PCR Forum


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 09-06-2006, 07:25 PM
moleculardude's Avatar
M.D/Ph.D
 
Join Date: Jul 2006
Location: Melbourne Australia
Posts: 423
Thanks: 5
Thanked 10 Times in 8 Posts
Default Primer Dimer problems in real-time PCR



I am having primer dimer problems in real-time PCR...

what tips do you guys have in fixing this. I even ordered the primers from a company they were supposed to be good!
Reply With Quote
  #2  
Old 11-18-2006, 09:56 PM
Pipette Filler
Points: 1,665, Level: 24 Points: 1,665, Level: 24 Points: 1,665, Level: 24
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2006
Posts: 7
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Primer Dimer problems in real-time PCR

I even use primer design software, what results Self Dimer dG
Mb u need to order/choose another primers?

Another way is:
- Set anneling time to minimun
- set concentration of primers to minimum
- increase Ta

or if u detect dimers only in low concentrations of substract u can try to add incubation b4 Plate Read just around 1-2 sec (Temperature: hi then T denaturation of dimers, but lower then T denaturation of ur product)

This information was taken form molbiol.ru
Sorry for my bad english
Reply With Quote
  #3  
Old 12-13-2006, 07:10 AM
Pipette Filler
Points: 1,562, Level: 23 Points: 1,562, Level: 23 Points: 1,562, Level: 23
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2006
Posts: 5
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Primer Dimer problems in real-time PCR

I first suggest that the primer concentration be set to a minimum after adjusting for optimal primer:target sequence ratio. I also suggest that one need to re-design the primers such that the last three NTDs at 3' end contain no G or Cs. Increasing the annealing temperature will not help much if the primer:target ratio is far from optimum
Reply With Quote
  #4  
Old 12-13-2006, 09:40 PM
kiki06's Avatar
Post-Doc
Points: 5,313, Level: 49 Points: 5,313, Level: 49 Points: 5,313, Level: 49
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2006
Posts: 277
Thanks: 0
Thanked 6 Times in 5 Posts
Default Re: Primer Dimer problems in real-time PCR

Yes, those are very good points.

I would ask for the sequences as mentioned and check them in a primer design program for self-annealing!

Reply With Quote
  #5  
Old 01-26-2009, 11:31 PM
Pipette Filler
Points: 48, Level: 1 Points: 48, Level: 1 Points: 48, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jan 2009
Posts: 9
Thanks: 2
Thanked 3 Times in 3 Posts
Default Re: Primer Dimer problems in real-time PCR

make a hot start and do not allow to stand your real time mix + template at room
temperature.
Reply With Quote
  #6  
Old 07-19-2010, 10:23 PM
Pipette Filler
Points: 78, Level: 1 Points: 78, Level: 1 Points: 78, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2010
Location: USA
Posts: 6
Thanks: 3
Thanked 2 Times in 2 Posts
Default Re: Primer Dimer problems in real-time PCR

Thanks
Reply With Quote
Reply

Tags
dimer , pcr , primer , problems , realtime


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
What is Gravity? sdr@sdrodrian.com Physics Forum 1 01-27-2008 02:53 PM
Moving Dimensions Theory--Returning Physical Models to Physics & Unifying QM & SR & GR & Statistical Mechanics Captain Ranger McCoy Physics Forum 8 08-29-2007 07:00 PM
Simply put, MOVING DIMENSIONS THEORY is THE NEW MODEL: http://physicsmathforums.com drelliot@gmail.com Physics Forum 0 08-29-2006 06:32 AM
The Theory of Moving Dimensions: Space and Time Are Moving Relative to One-Another Ranger West Physics Forum 8 01-14-2004 06:17 PM
The Law of Moving Dimenstions: Time Dimension is Moving Relative To Spatial Dimension Ranger West Physics Forum 6 11-12-2003 01:34 AM


All times are GMT. The time now is 09:20 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 0.17589 seconds with 16 queries