Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > General Science Forums > General Science Questions and Layperson Board
Register Search Today's Posts Mark Forums Read

General Science Questions and Layperson Board General Science Questions and Layperson Board


DNA digestion, ligation

DNA digestion, ligation - General Science Questions and Layperson Board

DNA digestion, ligation - General Science Questions and Layperson Board


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 11-02-2012, 08:23 PM
Pipette Filler
Points: 64, Level: 1 Points: 64, Level: 1 Points: 64, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2012
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default DNA digestion, ligation



Hi everyone,
I am facing problem at transformation step. After ligation I am not able to get the colonies. I change buffer, media and enzyme. Start getting colonies but those are not positive colonies. I have done digestion with Hind111 and XHO1. at 37 for 2hr then purified by qiagen gel extraction and ligate overnight at 4 oC.
My all grown positive colonies are showing smearing. What could be the reason. How I can troubleshoot this problem.
Reply With Quote
Reply

Tags
digestion , dna , ligation


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
DNA Ligation Problem Double Digestion Denise Molecular Cloning Forum 8 02-21-2012 06:40 AM
Problem in restrction digestion and ligation? srivels Molecular Cloning Forum 1 01-28-2010 05:02 PM
Ligation Deitiker, Philip R Protocols and Methods Forum 1 06-28-2007 12:40 PM
Ligation Deitiker, Philip R Protocols and Methods Forum 0 06-21-2007 11:11 PM
Ligation Deitiker, Philip R Protocols and Methods Forum 0 06-21-2007 09:12 PM


All times are GMT. The time now is 04:30 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 0.10878 seconds with 16 queries