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Micrococcal Nuclease

#EN0181	8000 u (300 u/µl)
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EN0181 MSDS (English) MSDS (English-USA) MSDS (German)

Description
Micrococcal Nuclease (S7 Nuclease) is a relatively nonspecific endo-exonuclease that digests single-stranded and double-stranded nucleic acids, but is more active on single-stranded substrates. Cleavage of DNA or RNA occurs preferentially at AT or AU-rich regions yielding mononucleotides and oligonucleotides with terminal 3'-phosphates.
The enzyme activity is strictly dependent on Ca²⁺.

Source
E.coli cells with a cloned nuc gene encoding Staphylococcus aureus extracellular nuclease (micrococcal nuclease).

Molecular Weight
16.9 kDa monomer.

Applications

• Hydrolysis of nucleic acids in crude cell-free extracts (1).
• Sequencing of RNA (2).
• Studies of chromatin structure (3).
• A model for protein folding and for structure-function studies (4, 5).

Quality Control
95% purity confirmed by SDS-PAGE.

Concentration
300 u/µl

Definition of Activity Unit
One unit of the enzyme releases 1.0 A₂₆₀ unit of acid-soluble products in 30 min at 37°C.
Enzyme activity is assayed in the following mixture: 100 mM sodium-glycine (pH 8.6), 10 mM CaCl₂ and 4.5 mM calf thymus DNA.

Storage Buffer
The enzyme is supplied in: 20 mM HEPES-KOH (pH 7.6), 50 mM NaCl and 50% (v/v) glycerol.

Inhibition and Inactivation

• Inhibitor: metal chelators, deoxythymidine 3'5'-diphosphate.
• Inactivated by the addition of EGTA or EDTA.

Related Product

Water, nuclease-free

References

1. Pelham, H.R.B., Jackson, R.J., An efficient mRNA-dependent translation system from reticulocyte lysates, Eur. J. Biochem., 67, 247-256, 1976.
2. Krupp, G., Gross, J.H., Rapid RNA sequencing: nucleases from Staphylococcus aureus and Neurospora crassa discriminate between uridine and cytidine, Nucleic Acids Res., 6, 3481-3490, 1979.
3. Telford, D.J., Stewart, B.W., Micrococcal nuclease: its specificity and use for chromatin analysis, Int. J. Biochem., 21, 127-137, 1989.
4. Tucker, P.W., et al., Staphylococcal nuclease reviewed: a prototypic study in contemporary enzymology. IV. The nuclease as a model for protein folding, Molec. Cell. Biochem., 23, 131-140, 1979.
5. Torchia, D.A., et al, Staphylococcal nuclease: sequential assignments and solution structure, Biochemistry, 27, 5509-5524, 1989.

Updated vasario 05, 2008 14:34