Thermo Scientific Micrococcal Nuclease (MNase) is suitable for removing nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments.
This micrococcal nuclease is a stable liquid form of the enzyme derived from Staphylococcus aureus. Micrococcal nuclease (MNase) exhibits exo- and endo-5'-phosphodiesterase activities against DNA and RNA. This enzyme digests double-stranded, single-stranded, circular and linear nucleic acids. The highest activity is toward single-stranded nucleic acid substrates with preference for AT- or AU-rich regions. Enzymatic activity occurs at pH 7 to 10 and is strictly dependent on calcium for digestion of RNA and DNA substrates. Micrococcal Nuclease is suitable for removing nucleic acids from cell lysates, releasing chromatin-bound proteins and shearing chromatin for use in chromatin immunoprecipitation (ChIP) experiments.
- Enzyme digests nucleic acids (DNA and RNA)
- Effective for double-stranded, single-stranded, circular and linear nucleic acids
- Active in neutral to alkaline buffer conditions containing divalent calcium ions
- Supplied at ≥100 units/µL in 10mM Tris-HCl pH 7.5, 50mM NaCl, 1mM EDTA, 50% glycerol
Properties of Micrococcal Nuclease (MNase):
- Product is a liquid at -20°C. Keep enzyme on ice during use.
- Micrococcal nuclease is dependent on Ca++ for activity. Avoid calcium chelators, such as EGTA, in reaction buffers.
- Enzyme is active at pH 7 to 10 with a salt concentration less than 100mM. Optimal enzyme activity occurs at 37°C; however, the enzyme is active at room temperature. Monovalent metal ions, such as Na+ and K+, will decrease activity. EGTA or heating to 65°C for 10 minutes will inactivate the enzyme.
Generalized Protocol for use of Micrococcal Nuclease (MNase):
- Dilute sample in 50mM Tris-HCl pH 8.0, 5mM CaCl2
- If sample only contains nucleotides (i.e., no protein), add BSA at a final concentration of 0.1mg/mL.
- Add micrococcal nuclease and incubate reaction at 37°C until the nucleotides are degraded.
- Optional: Stop reaction by adding EGTA, pH 8.0 to a final concentration of 20mM.
Specifications of Micrococcal Nuclease (Part No. 88216):
- Visual: Clear, colorless liquid free of insoluble material.
- Units: ≥100 units/µL
- Additional Information: 1 unit is defined as the amount of enzyme required to produce an increase in absorbance at 260nm of 0.001/min/mL at 25°C of highly polymerized DNA.
Review of chromatin IP (ChIP) assays
Pierce ChIP Kit
Subcellular Fractionation Kit