3 replies to this topic

[Rsm]

Hi there,
I would like to perform a ChIP-seq on primary cells (FACS sorted). However, I can get only a limited number of cells per mouse (~3000/mouse). So I'd end up with 20,000 cells, is that enough for ChIP-seq? How about reproducibility? I read the paper from the Myers' lab, they used 50ng DNA and amplified it. 20,000 cells should give me ~200ng DNA, that might work. Has anyone experience with small cell numbers for ChIP-seq? Is there a protocol or article describing ChIP-seq on small number of sorted cells?
Thanks a lot!
Cheers,
Minna

[TracyDuke]

I have never tried ChIPing using less than 1 million cells but you can check out this paper: Genome-Scale ChIP-chip analysis using 10,000 human cells. Luis G. Acevedo et al. http://www.pubmedcentral.nih.gov/articlere...bmedid=18251256

[cellcounter]

TracyDuke, on Aug 17 2009, 02:58 PM, said:

I have never tried ChIPing using less than 1 million cells but you can check out this paper: Genome-Scale ChIP-chip analysis using 10,000 human cells. Luis G. Acevedo et al. http://www.pubmedcentral.nih.gov/articlere...bmedid=18251256

Good paper! Thanks. I was also thinking of doing Chip-seq on limited supply of cells.

[Rsm]

Excellent paper!
That was exactly what I was looking for!
(even though they are using cultured cells )...
Thanks a lot!

Cheers,
Minna