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!
- - - - -

Input reverse cross linking

input reverse cross link

  • Please log in to reply
1 reply to this topic

#1 oxygene8



  • Members
  • Pip
  • 2 posts

Posted 21 April 2012 - 03:25 AM

I am performing a ChIP assay using the R&D kit. According to the manufacturers protocol, the samples (antibody IP and IgG IP) are washed after the antibody incubation and then boiled with a Chelating Resin solution for 10 minutes. This basically is the final step before DNA purification using a kit. I am assuming this boiling step involves reverse cross linking for the samples. My question is, how do I reverse cross link the input DNA that I have set aside previously? The kit nowhere says anything about the processing of input DNA. Can I add Chelating Resin solution to the input and treat likewise or do I follow a traditional 65 degrees treatment (though I am reluctant to use different methods for the input and samples).

Any help or experience with a such a problem will be welcome....




  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 154 posts

Posted 21 April 2012 - 02:18 PM

In my experience, boiling in either chelex or high pH Tris-EDTA buffers (they're essentially equivalent) is sufficient for reversal of crosslinking of input samples.

Edited by KPDE, 21 April 2012 - 02:19 PM.

Also tagged with one or more of these keywords: input, reverse cross link

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

©1999-2013 Protocol Online, All rights reserved.