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

Molecular Cloning Forum DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.


Ligated DNA to Competent E.Coli Cells for Midi Prep?

Ligated DNA to Competent E.Coli Cells for Midi Prep? - Molecular Cloning Forum

Ligated DNA to Competent E.Coli Cells for Midi Prep? - DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 12-20-2008, 01:42 AM
Pipette Filler
Points: 72, Level: 1 Points: 72, Level: 1 Points: 72, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2008
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Question Ligated DNA to Competent E.Coli Cells for Midi Prep?



Hi every body,
I face one experimental problem....I did ligation and now I want to transfer this ligated DNA ( Recombinanted Plasmide ) to competent E.Coli cells by electroporation so that I can get more desire recombinant plasmide by midiprep...But somehow It dose not work...If some one have any idea how to proceed...Thanks...
Reply With Quote
  #2  
Old 12-20-2008, 08:05 AM
butters's Avatar
M.D/Ph.D
Points: 4,938, Level: 47 Points: 4,938, Level: 47 Points: 4,938, Level: 47
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2008
Location: Malaysia
Posts: 315
Thanks: 25
Thanked 68 Times in 63 Posts
Default Re: Ligated DNA to Competent E.Coli Cells for Midi Prep?

uh u tried the calcium chloride method?
Reply With Quote
  #3  
Old 02-26-2009, 01:37 PM
Pipette Filler
Points: 3,020, Level: 35 Points: 3,020, Level: 35 Points: 3,020, Level: 35
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Apr 2006
Location: Bavaria, Germany
Posts: 18
Thanks: 0
Thanked 4 Times in 3 Posts
Arrow Re: Ligated DNA to Competent E.Coli Cells for Midi Prep?

Did you remove the salt before electroporation?

Quote:
Originally Posted by sunrise View Post
Hi every body,
I face one experimental problem....I did ligation and now I want to transfer this ligated DNA ( Recombinanted Plasmide ) to competent E.Coli cells by electroporation so that I can get more desire recombinant plasmide by midiprep...But somehow It dose not work...If some one have any idea how to proceed...Thanks...
Reply With Quote
Reply

Tags
cells , competent , dna , ecoli , ligated , midi , prep


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
mouse hepatocytes isolation amicia Cell Biology and Cell Culture 39 05-11-2012 11:52 AM
Characteristics of cancerous cells aftabac Cell Biology and Cell Culture 2 08-26-2011 11:40 PM
Human Cytome Project - Update 24 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 1 08-01-2010 02:18 PM
How to isolate adult from circulation? peterish Stem Cell Forum 0 05-19-2008 10:02 PM
Human Cytome Project - Update 6 Jan. 2005 Peter Van Osta Cell Biology and Cell Culture 0 01-06-2005 11:18 AM


All times are GMT. The time now is 05:51 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 0.15650 seconds with 16 queries