Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Basic Lab Protocols and Techniques
Register Search Today's Posts Mark Forums Read


Annealing Issues

Annealing Issues - Basic Lab Protocols and Techniques

Annealing Issues -


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 06-15-2010, 06:29 PM
Pipette Filler
Points: 12, Level: 1 Points: 12, Level: 1 Points: 12, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2010
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Annealing Issues



I am having trouble with an annealing reaction where I need to clean the single stranded oligos from the tube containing the double stranded product as the oligos would interfere with the downstream parts of my protocol. Are there any suggestions as to how to do this?

I have tried running an agarose gel, which produces two bands. I have thought about cutting out the larger band, but this is complicated by the fact that single stranded DNA does not migrate in a predictable fashion on a gel.

Any advice would be appreciated.
Reply With Quote
  #2  
Old 06-15-2010, 09:27 PM
Graduate Student
Points: 2,626, Level: 33 Points: 2,626, Level: 33 Points: 2,626, Level: 33
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2010
Location: Santiago, Chile
Posts: 100
Thanks: 1
Thanked 24 Times in 24 Posts
Default Re: Annealing Issues

I don't think it'd be a problem cutting your target DNA band, and I think you won't have any oligos contaminating your sample, because oligos always run in the front of the gel, either lineal or as dimers, 'cause they're so small.
Reply With Quote
  #3  
Old 07-08-2010, 01:59 PM
Pipette Filler
Points: 3,433, Level: 38 Points: 3,433, Level: 38 Points: 3,433, Level: 38
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2007
Posts: 13
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Annealing Issues

Hai ,
I too agree with prevoius reply. just run a normmal agarose gel. your ssDNA oligo should run out of the gel or show at the bottom as it runs faster than dsDNA.
Reply With Quote
Reply

Tags
annealing , issues


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
Annealing temp using Quikchange SDM kit Clarisa Bejar Protocols and Methods Forum 2 03-22-2008 04:13 PM
Annealing Temperatures for PCR Jayakumar, R Protocols and Methods Forum 0 03-30-2006 02:34 PM
Call for IPSI Transactions Special Issues in 2005/6; c/ba IPSI Transactions Special Issues Protein Forum 0 02-26-2005 12:48 AM
FA: Physics Today, issues Oct 73 to May 86 & more Dan Physics Forum 0 12-27-2004 05:05 PM
Ligase free annealing question EK Protocols and Methods Forum 9 01-24-2004 11:15 AM


All times are GMT. The time now is 07:41 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12195 seconds with 16 queries