Go Back   Molecular Biology Forum > Molecular Research Topics Forum > PCR - Polymerase Chain Reaction Forum > Real-Time PCR and Quantitative PCR Forum
Register Blogs FAQ Members List Calendar Science Groups New! Arcade Search Today's Posts Mark Forums Read

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


real-time Videos
real-time-pcr-protocol-of-gm-soybean-dna
real-time-pcr-protocol-of-gm-soybean-dna Real-Time PCR Protocol of GM Soybean DNA. From the Youtube user tangodurden.


Primer Dimer problems in real-time PCR

Real-Time PCR and Quantitative PCR Forum

Real-Time PCR and Quantitative PCR Forum



Register Molecular Biology Forums
Reply
 
LinkBack Thread Tools Display Modes
  #1 (permalink)  
Old 09-06-2006, 07:25 PM
moleculardude's Avatar
M.D/Ph.D
Points: 5,094, Level: 48Points: 5,094, Level: 48Points: 5,094, Level: 48
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Jul 2006
Location: Melbourne Australia
Posts: 304
Blog Entries: 1
moleculardude RSS Feed
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!
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Alt Today
Advertising
Google Adsense
 
This advertising will not be shown
in this way to registered members.
Register your free account today
and become a member on
Molecular Biology Forum
Standard Sponsored Links

  #2 (permalink)  
Old 11-18-2006, 08:56 PM
Pipette Filler
Points: 1,062, Level: 18Points: 1,062, Level: 18Points: 1,062, Level: 18
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Nov 2006
Posts: 7
Leowe RSS Feed
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
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #3 (permalink)  
Old 12-13-2006, 06:10 AM
Pipette Filler
Points: 990, Level: 17Points: 990, Level: 17Points: 990, Level: 17
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Dec 2006
Posts: 5
waleedkars RSS Feed
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
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #4 (permalink)  
Old 12-13-2006, 08:40 PM
kiki06's Avatar
Post-Doc
Points: 4,288, Level: 44Points: 4,288, Level: 44Points: 4,288, Level: 44
Activity: 0%Activity: 0%Activity: 0%
 
Join Date: Feb 2006
Posts: 263
kiki06 RSS Feed
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!

Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
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
Real time PCR - neophyte .. indigo_nation Real-Time PCR and Quantitative PCR Forum 1 02-08-2008 08:41 PM
How to get rid of PCR primer dimers molecule2005 PCR - Polymerase Chain Reaction Forum 0 11-09-2007 02:04 PM
GSP Gene Specific Primers for Real Time PCR dnamolecule Real-Time PCR and Quantitative PCR Forum 1 10-03-2007 04:45 PM
BSA Albumin for Real Time PCR? pipette_man Real-Time PCR and Quantitative PCR Forum 1 10-03-2007 04:37 PM
Real Time PCR Machines Thermocyclers MARGARITA MARTINEZ Lab Equipment Discussion and Reviews 4 07-29-2007 09:52 PM


All times are GMT. The time now is 02:02 PM.


Powered by vBulletin® Version 3.7.1
Copyright ©2000 - 2008, Jelsoft Enterprises Ltd.
Copyright 2005-2007 Molecular Station | All Rights Reserved