Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

Concentration of DNA for Transformation

  • Please log in to reply
1 reply to this topic

#1 DNA



  • Active Members
  • PipPipPipPipPip
  • 42 posts

Posted 24 October 2009 - 01:43 AM

Hi All,

What should be teh optimum concentration of plasmid DNA in order to have successful transformation and get well isolated colonies.
I have been facing problems in determining the amount of DNA needed for transformation and most of the time i end up with smear on the plates rather than well separated colonies so I then take some smear from that and re-plate them and so on....

#2 phage434



  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,747 posts

Posted 24 October 2009 - 06:39 AM

If you are transforming miniprep plasmid, then 100 pg of plasmid will yield hundreds or thousands of transformants, assuming you have good transformation efficiency. Usually people plate two or more plates, with different amounts or dilutions of the transformed cells, using the one with the isolated colonies. Are you sure your antibiotic selection is working? Do you plate untranformed cells to check?

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.