Source: Aspergillus oryzae
Non-specific endonuclease, active primarily on single-stranded DNA and RNA.
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Catalyzes degradation of single-stranded nucleic acids into oligonucleotides and 5’-mononucleotides (1).
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Cleaves both single-stranded DNA and RNA, with stronger DNAse acitivity.
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Double-stranded DNA, double-stranded RNA and DNA-RNA hybrids are resistant to degradation at moderate enzyme concentration.
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Capable of cleavage of double-stranded nucleic acids at nicks, mismatches and small gaps (2).
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Relaxes/linearizes supercoiled plasmids.
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Removes protruding single-stranded ends.
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Used for S1 mapping of nucleic acids.
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Requires Zn2+ ions for activity.
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The enzyme is active up to 65°C.
Storage Buffer: 20 mM Tris-HCl (pH 7.5 at 22°C), 50 mM NaCl, 0.1 mM ZnCl2 and 50% (v/v) glycerol.
Assay Conditions: 30 mM sodium acetate (pH 4.6), 1 mM zinc acetate, 50 mM NaCl, 0.5 mg/ml of activated DNA, 5% glycerol. Incubation is at 37°C for 10 min in a reaction volume of 0.5 ml.
Quality Control: All preparations are assayed for contaminating exonuclease and double-stranded DNase activities.
References:
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Berg, A.J. et al. (1977) Cell 12, 721.
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Sambrook, J. et al. (1989) Molecular cloning: A laboratory Manual, second edition, pp. 5.78-5.79, Cold Spring Harbor, New York.