Start working on ChIP - any suggestion for the novice? (Oct/13/2005 )

Dear All,

I am going to do chromatin CHIP experiment soon. I heard that this method is not so easy to do
Can anyone give me any suggestion about what kits I should use, and any suggestion about this method? I am suggested to use MNase instead of sonication for chromatin CHIP.

I will also appreciate if someone can kindly help me to get a paper about this method:
Umlauf D, Goto Y, Feil R.
Site-specific analysis of histone methylation and acetylation.
Methods Mol Biol. 2004;287:99-120. Review.

Million thanks for any help.

Best Wishes
Hy

Most people use the kit or similar conditions to the kit sold by upstate biosciences. As far as MNase treatment goes, it is more difficult but theoretically can produce cleaner results. Most people use sonication to sheer the DNA, just make sure that you do the controls to work out the conditions to use and run it out on a gel to visualize the size of the fragments that you obtain. In most cases people want <1000bp

I have just started doing ChIP but I am learning from so serious experts. They swear by their own non kit based method and in my novice hands it's producing good results. The method uses MNase digestion not sonication and can be X-linked or native dependant on what you want to look at and the resolution required.

Wow, you are such a lucky guy... So it sounds MNase has more advantage than sonication?
If you do not mind, may I share your protocol? Many thanks