- Digestion of nucleotides included in crude cell extract
- Digestion of chromatin for nucleosome preparation
- Chromatin immunoprecipitation (ChIP)
10X Micrococcal Nuclease [200 mM Tris-HCl (pH 8.0), 50 mM NaCl, 25 mM CaCl2]
Nuclease activity is not detected after incubation of 1 µg lambda DNA with 50 units of this enzyme for 10 minutes at 37°C in the reaction enzyme buffer (1 x H buffer), as judged from the agarose gel electrophoresis pattern. This enzyme is more than 95% homogenous as judged by SDS-PAGE.
1. Lefevre, P. & Bonifer, C. (2006) Methods Mol. Biol. 325:315-325.
2. Heins, J.N. et al. (1966) Procedures Nucl. Acids Res. 1:79.
3. Pelham, H.R. & Jackson, R.J. (1976) Eur. J. Biochem. 67:247-256.
4. Telford, D.J. & Stewart, B.W. (1989) Int. J. Biochem. 21:127-137.
One unit is defined as the amount of enzyme required to produce 1 OD260 of acid-soluble product in 30 minutes at 37°C and pH 8.0 using heat-denatured calf thymus DNA as the substrate.
50 mM Tris-HCl (pH 8.0), 10 mM 2-mercaptoethanol, 50 mM NaCl, 50% glycerol