How to do good DNaseI Hypersensitivity Sites Assay?

h9290184 at hkucc.hku.hk h9290184 at hkucc.hku.hk
Sat Feb 19 01:44:54 EST 1994


Dear Netters,
	I'm currently working on DNaesI Hypersensitivity Sites assay, but after 
partial DNaseI digestion, I find it very difficult to obtain a complete
restriction enzyme digestion (usually I need to digest several tens of ug of 
DNA if not more). The partially digested DNA usually gives obscure result
during intrepretation esp. when looking at the hyper-sites of high molecular
weight (>5kb). I have tried repeated protease K digestions on my sample as well 
as repeated and prolong restriction enzyme (10X enzyme, three times, >24hrs),
spermidine was also added during the digestion but the situation wasn't
improved. Is there anybody out there has a good protocol for this assay or
any suggestion of improving the restriction enzyme digestion? 
	Zillion thanks!

Theobald Chan
PS. I am now using a borrowed account, email answer here or post will be fine.



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