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Old 04-06-2011, 01:02 PM   #1
neocortex
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Question ChIP-seq with very small amount of tissue

Hi,

We are trying to do ChIP with very small amount of tissue, like <10mg, which is tightly limited by the technical difficulty in collecting the tissue. Does anyone has experience doing ChIP with this little tissue, and obtained enough DNA for sequencing? If you would like to share some of your experience, or give me some advice, I would really appreciate it!! Thanks a lot!!
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Old 04-07-2011, 04:25 AM   #2
mudshark
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Did you already read this?

Nature Genetics 38, 835 - 841 (2006)
Epigenetic characterization of the early embryo with a chromatin immunoprecipitation protocol applicable to small cell populations

Laura P O'Neill1, 2, Matthew D VerMilyea1, 2 & Bryan M Turner

http://www.nature.com/ng/journal/v38/n7/abs/ng1820.html

and

http://www.epigenome-noe.net/WWW/res....php?protid=26


I guess that after ChIP you will have to do a serious amplification of your IP material using WGA e.g.

Last edited by mudshark; 04-07-2011 at 04:31 AM.
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Old 04-11-2011, 11:13 AM   #3
neocortex
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Hi,

Thanks for the info. Sorry maybe I did not make it clear, that the samples I deal with are actually pieces of tissues from microdissection and I had to freeze it down immediately after the dissection.

The paper linked above was using freshly isolated tissue/cells. It's really different. I am just wondering what would be the best way to homogenize, how much tissue to start with and how much chromatin you can get from it.

Thanks,
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Old 04-13-2011, 07:06 AM   #4
mudshark
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Sorry. I have no experience with ChIP from tissues at all.

maybe the following link can help you:
http://www.abcam.com/index.html?page...urce&rid=11803

you also could check out ChIP kits offered by various companies (Diagenode, Millipore etc)
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