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!
- - - - -

isolating whole chromatin before ChIP

  • Please log in to reply
1 reply to this topic

#1 crockel



  • Active Members
  • Pip
  • 8 posts

Posted 07 December 2012 - 03:51 AM

Dear all,

I am performing ChIPseq on overexpressed TFs in Drosophila wing discs. After x-linking (1% FA, 10 min) I have the problem that a two step lysis does not work anymore. Basically I am only disrupting the wing discs and lysing the cells during sonication. One of my presumed problems is now, that there might be a lot of unbound (cytosolic) TF, which I precipitate and I want to decrease this precipitation of "useless" TF. Therefore I thought to precipitate/isolate the whole chromatin before I do the ChIP. Does anyone know a good strategy how to do this? Would using spermidine do the job?

Thanks for your help,

#2 pcrman



  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts

Posted 07 December 2012 - 09:00 AM

You may try the method described in this paper to isolation chromatin fraction from fly http://www.nature.co...ature10492.html

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

©1999-2013 Protocol Online, All rights reserved.