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
- - - - -

Vector Fill in for Blunt End Cloning


  • Please log in to reply
No replies to this topic

#1 Euclid

Euclid

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 24 March 2009 - 08:14 AM

I have a blunt ended insert I need to clone into a vector that was cut with a sticky end restriction enzyme. Using a blunt end enzyme to cut the vector is not an option. I cut the vector and filled in with klenow. Do I need to treat the linear, filled in vector with CIP? I do not think there are phosphates on the ends after fill in, because when I do this with to an insert I need to add phosphates with PNK.

Anybody have experience with this technique and able to share it?

Thanks!




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

©1999-2013 Protocol Online, All rights reserved.