Micrococcal Nuclease

Micrococcal Nuclease is an endonuclease that preferentially digests single-stranded DNA or RNA, especially at AT- or AU-rich regions. The enzyme will also digest double-stranded DNA or RNA, making it an essential component of chromatin immunoprecipitation (ChIP) assays. Micrococcal Nuclease digests 5'-phosphodiester bonds of DNA and RNA, yielding 3'-phosphate mononucleotides and oligonucleotides. This enzyme requires Ca2+ as a cofactor for its activity and is completely inactivitated by EDTA or EGTA.

Applications

Digestion of nucleotides included in crude cell extract
Digestion of chromatin for nucleosome preparation
Chromatin immunoprecipitation (ChIP)

Supplied buffer

10X Micrococcal Nuclease [200 mM Tris-HCl (pH 8.0), 50 mM NaCl, 25 mM CaCl2]

Purity

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 (1X H buffer), as judged from the agarose gel electrophoresis pattern. This enzyme is more than 95% homogenous as judged by SDS-PAGE.

Form

50 mM Tris-HCl (pH 8.0), 10 mM 2-mercaptoethanol, 50 mM NaCl, 50% glycerol

Product citations

Lefevre, P. & Bonifer, C. Analyzing Histone Modification Using Crosslinked Chromatin Treated With Micrococcal Nuclease. Methods Mol. Biol. 325:315–325 (2006).

Heins, J.N. et al. Procedures Nucl. Acids Res. 1:79 (1966).

Pelham, H.R. & Jackson, R.J. An Efficient mRNA-Dependent Translation System from Reticulocyte Lysates. Eur. J. Biochem. 67:247–256 (1976).

Telford, D.J. & Stewart, B.W. Micrococcal nuclease: Its specificity and use for chromatin analysis. Int. J. Biochem. 21:127–137 (1989).