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
- - - - -

make a Mega plasmid mutation


  • Please log in to reply
1 reply to this topic

#1 summersnows

summersnows

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 17 December 2010 - 12:50 AM

Hi there.

I'm study a Mega plasmid;220kb from Environmental strain.

in this case, i want make random mutation or point mutation.

how can i do that?

it's size make me crazy.

#2 bsksln

bsksln

    member

  • Active Members
  • Pip
  • 11 posts
0
Neutral

Posted 17 December 2010 - 10:04 PM

some ecoli strains contain lambda phage red recombination system. you can design two homology arms around your mutation site and transform it into this ecoli strain containing your huge plasmid. typically the mutation can be introduced into the plasmid by recombination. in our lab we often use this strain to construct vectors.
however there are many practical problem using this method. you can send me message if you are considering to use this method.




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

©1999-2013 Protocol Online, All rights reserved.