Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum

Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum (http://www.molecularstation.com/forum/)
-   DNA Extraction Forum (http://www.molecularstation.com/forum/dna-extraction-forum/)
-   -   ChIP Preclear Step (http://www.molecularstation.com/forum/dna-extraction-forum/73135-chip-preclear-step.html)

BakedGoat 06-10-2010 06:31 PM

ChIP Preclear Step
 
Hi everyone,

I'm fairly new to ChIP and am working through my first run at the protocol. Basically, when I was running through the preclear step, I was incubating for 30 minutes, ran out for a coffee, and when I got back, I added my antibody to my samples without first spinning down and removing the supernatant. So I ended up incubating all night with my agarose beads, and antibody in my DNA sample. I was wondering how crucial it is to remove the beads?

Pippuri 06-21-2010 11:02 PM

Re: ChIP Preclear Step
 
Hi BakedGoat,

It is ESSENTIAL to remove the beads indeed in order to achieve the purpose of that particular step: preclearing. As the purpose of this step is to remove all the nonspecific stuffs that will interference with your later step. Without removing the beads, all the nonspecific stuffs will be carried over, and you end up getting pretty dirty pull down...

Hope this helps.


All times are GMT. The time now is 05:48 AM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved

Page generated in 0.09046 seconds with 11 queries