how to purify ChIP DNA by using kit - (Aug/18/2007 )

Hello,
I will try to use Qiagen Kit for ChIP DNA purification. But I do not know which kit should I use or at which step should I use the buffers of the kit?
If anyone has any exyperience, would be wonderful to hear.
Thanks

-ehallacli-

you can use the qiagen PCR purification kit.

-pcrman-

QUOTE (pcrman @ Aug 19 2007, 05:44 PM)

you can use the qiagen PCR purification kit.

So after decrosslinking, you just apply binding buffer? Or do you also make deproteination by Proteinase K?
I could not figure out

Thanks

-ehallacli-

QUOTE (ehallacli @ Aug 20 2007, 05:32 AM)

QUOTE (pcrman @ Aug 19 2007, 05:44 PM)

you can use the qiagen PCR purification kit.

So after decrosslinking, you just apply binding buffer? Or do you also make deproteination by Proteinase K?
I could not figure out

Thanks

Hi,

I presume you must have used protein G (or A) agaore and eluted DNA from beads before you do decrosslinking then add it to spin column, right?

Well, proteinase K is an essential step, before you can really purify chromatin DNA from column. I experienced lower yeild of DNA and poor sonication result owing to a bad quality proteinase K.

-freedomal-

QUOTE (freedomal @ Aug 20 2007, 04:34 AM)

QUOTE (ehallacli @ Aug 20 2007, 05:32 AM)

QUOTE (pcrman @ Aug 19 2007, 05:44 PM)

you can use the qiagen PCR purification kit.

So after decrosslinking, you just apply binding buffer? Or do you also make deproteination by Proteinase K?
I could not figure out

Thanks

yes. My protein is TAP tagged so i am using IgG beads anb eluting them with 1% SDS in 65 degress.
so I will decrosslink, then ProtK and qiagens PB buffer

Thank you very much for the tips.

-ehallacli-