S1 Nuclease

S1 Nuclease is a non-specific endonuclease, active primarily on single-stranded DNA and RNA.

Source: Aspergillus oryzae

Description: 

· Catalyzes degradation of single-stranded nucleic acids into oligonucleotides and 5’-mononucleotides (1).

· Cleaves both single-stranded DNA and RNA, with stronger DNAse acitivity.

· Double-stranded DNA, double-stranded RNA and DNA-RNA hybrids are resistant to degradation at moderate enzyme concentration.

· Capable of cleavage of double-stranded nucleic acids at nicks, mismatches and small gaps (2).

· Relaxes/linearizes supercoiled plasmids.

· Removes protruding single-stranded ends.

· Used for S1 mapping of nucleic acids.

· Requires Zn²⁺ ions for activity.

· 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:

1. Berg, A.J. et al. (1977) Cell 12, 721.

2. Sambrook, J. et al. (1989) Molecular cloning: A laboratory Manual, second edition, pp. 5.78-5.79, Cold Spring Harbor, New York.