Single-stranded specific DNA and RNA endonuclease.
· 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 37oC using denatured calf thymus DNA.
Storage Conditions: Store at -20oC.
Storage Buffer: 10 mM Tris-HCl (pH 7.5 at 22oC), 0.1 mM zinc acetate, and 50% (v/v) glycerol.
Reaction Buffer: 30 mM sodium acetate (pH 5.0 at 22oC), 30 mM NaCl, 1 mM ZnCl2.
Note: Also active in Low & Medium Buffers.
Assay Conditions: 30 mM sodium acetate (pH 5.0 at 22oC), 50 mM NaCl, 1 mM ZnCl2, 0.5 mg/ml denatured calf thymus DNA and 5% glycerol. Incubation is at 37oC for 10 min in a reaction volume of 1 ml.
Quality Control: All preparations are assayed for contaminating double-stranded DNase activity.
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.