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Epigenetics Forum: DNA Methylation, Histone and Chromatin Study An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin


Nuclear Isolation ChIP Chromatin IP

Epigenetics Forum: DNA Methylation, Histone and Chromatin Study

An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin



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Old 02-14-2008, 03:33 PM
Pipette Filler
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Question Nuclear Isolation ChIP Chromatin IP

Hi I am trying to isolate nuclei to conduct a ChIP assay.

Does anyone have experience to do this?

I am using 293 cells transfected with my plasmid.

Do you guys use Triton-X 100 or NP-40 detergents?

cheers
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  #2 (permalink)  
Old 03-07-2008, 10:39 PM
Pipette Filler
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Default Re: Nuclear Isolation ChIP Chromatin IP

the protocols out there use Triton X-100 for some buffers and NP-40 for others. The cell lysis buffer is with 0.5% NP-40. Then the nuclei are suspended in a lysis buffer with 1% SDS. For the washing buffers : the Low and high salt are with 0.1% SDS and 1% triton X-100 . The LiCl wash buffer is with 1% NP40. You can find one of these protocols in Millipore website.
Good luck
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