ChIP from E7.5 and E8.5 Mouse Embryos

[utkarshkapoor]

Hi,

The aim of my project is to carry out chromatin immuno-precipitation for a protein that would probably be present in about 5-10% cells of the entire embryo. Since the size of the embryo is small, I don't have an estimate of the amount of starting material I should use for the exercise. Any suggestions are welcome.

People who have experience in doing ChIP from mouse embryos, please help out!

Thanks,

Utkarsh

[jwfoley]

A lot of people might either grow a cell culture or pool multiple individuals, though those both have major downsides.

There's a published protocol using the cumbersome T7 method for linear amplification from small samples: doi:10.1038/nmeth.1626. I've heard this is difficult. Or you can apparently use NuGEN's very slick SPIA method, which would be much simpler, but I haven't seen it published: http://www.nugen.com/nugen/index.cfm...lication-note/.