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ChIP Preclear Step
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?
Re: ChIP Preclear Step
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.
|chip , preclear , step|
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