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
- - - - -

plasmid + sepharose


  • Please log in to reply
1 reply to this topic

#1 sorcener

sorcener

    member

  • Active Members
  • Pip
  • 13 posts
0
Neutral

Posted 30 June 2010 - 02:54 PM

Hi to all,

I had normally made maxipreps using the super easy kits protocols (i.e. Qyagen, Invitrogen...) but i have arrived to a lab were they make the maxipreps in a old way using home made solutions ans sepharose columns.


Briefly, the procedure involves an alkaline lisis, clearing, RNAse A incubation, Phenol-cloroform extraction and passing the resulting suppernatant to the sepahrose column for recovering several fractions, some of them containing the plasmid.
I did it once but got really few plasmid!!!

My question is, if somebody still do it in this way, which is the function of the sepharose column? it it really neccesary after phenol cloroform extraction?

thanks inadvance

#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,817 posts
137
Excellent

Posted 01 July 2010 - 06:17 AM

the sepharose column is to separate the molecules by size. in your case, separating plasmid from genomic dna.
talent does what it can
genius does what it must
i do what i get paid to do




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

©1999-2013 Protocol Online, All rights reserved.