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!
Photo
- - - - -

Ligation with PLAFR


  • Please log in to reply
1 reply to this topic

#1 alshia

alshia

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 09 June 2004 - 07:12 PM

Please help
I am trying to ligate fragments of DNA that have been double digested with Bgl II and Hind III into the vector PLAFR3.
I am making my cells competent by washing in Calcium chloride and using freeze thaw to transform them.
can anyone offer sugesstions of why this is not working :blink:

#2 pcrman

pcrman

    Epigenetist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts
68
Excellent

Posted 09 June 2004 - 07:56 PM

Hi alshia,

Many things in your experiment could have gone wrong, such as ligation, preparation of competent cells, and transformation. Before others could answer your question, please provide some details:

What type of your DNA is, PCR product or DNA cut from from a vector?
How did you prepare your vector?




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

©1999-2013 Protocol Online, All rights reserved.