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