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Fast ChIP + purification column


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#1 Radish

Radish

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Posted 19 November 2009 - 05:32 PM

Hello to you all!

I am a long time fan of your forum, so I finally decided to get out of the bushes, stop lurking and actually participate in the forum discussions  :D .

I have been reading for a long time the advantages of the fast chip protocol when compared to the regular protocols, so I finally decided to put it in practice. However I am a little confused, do I need to do the column purification (I read in the forum arquives that you use a qiagen column to purify the DNA instead of doing the phenol chlorform extraction)? And if so, which type do you use?

To clarify my doubt:

1) crosslink and harvest cells
2)lyse cells and wash pellet
3)shear chromatine and centrifuge
4)add ab
5)centrifuge to clear aggregates
6)add protein A
7)wash beads
8)add chelex and boil
9)add proteinase K and incubate
10) boil
11) centrifuge and collect supernatant
12) column purification? :lol:

Thank you so much to all of you. Your help will be greatly appreciated.
Oh, and thank you jiro_killua I learned to do the ChIP data analysis from you  :P




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