Mung Bean Nuclease

Single-stranded specific DNA and RNA endonuclease.

Description:

· Removes protruding ends in duplex DNA resulting in ligatable blunt ends (1).

· Suitable for trimming single-stranded protruding ends of DNA or RNA without degrading the duplex structure (2).

· Used for mapping of transcripts by analyzing the Mung Bean Nuclease-resistant RNA-DNA hybrids (3).

· Digests hairpin structures during cDNA synthesis (4).

· Will not cleave the strand opposite a nick in duplex DNA.

· Requires Zn²⁺ ions for activity.

Unit Definition: One unit is the amount of enzyme required to produce 1 µg of acid-soluble material per minute at 37^oC using denatured calf thymus DNA.

Storage Conditions: Store at -20^oC.

Storage Buffer: 10 mM Tris-HCl (pH 7.5 at 22^oC), 0.1 mM zinc acetate, and 50% (v/v) glycerol.

Reaction Buffer: 30 mM sodium acetate (pH 5.0 at 22^oC), 30 mM NaCl, 1 mM ZnCl₂.

Note: Also active in Low & Medium Buffers.

Assay Conditions: 30 mM sodium acetate (pH 5.0 at 22^oC), 50 mM NaCl, 1 mM ZnCl₂, 0.5 mg/ml denatured calf thymus DNA and 5% glycerol. Incubation is at 37^oC for 10 min in a reaction volume of 1 ml.

Quality Control: All preparations are assayed for contaminating double-stranded DNase activity.

References:

1. Ardelt, W., and Laskowski, M., Sr. (1971) Biochem. Biophys. Res. Commun. 44, No. 5, 1205-1211.

2. Berk, A.J., and Sharp, P.A. (1977) Cell 12, 721-732.

3. Berk, A.J., and Sharp, P.A. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 1274-1278.

4. Goodman, H.M. and McDonald, R.J. (1979) Methods Enzymol. 68, 75-90.