Single-stranded specific DNA and RNA endonuclease.
Description:
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Removes protruding ends in duplex DNA resulting in ligatable blunt ends (1).
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Suitable for trimming single-stranded protruding ends of DNA or RNA without degrading the duplex structure (2).
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Used for mapping of transcripts by analyzing the Mung Bean Nuclease-resistant RNA-DNA hybrids (3).
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Digests hairpin structures during cDNA synthesis (4).
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Will not cleave the strand opposite a nick in duplex DNA.
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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°C using denatured calf thymus DNA.
Storage Conditions: Store at -20°C.
Storage Buffer: 10 mM Tris-HCl (pH 7.5 at 22°C), 0.1 mM zinc acetate, and 50% (v/v) glycerol.
Reaction Buffer: 30 mM sodium acetate (pH 5.0 at 22°C), 30 mM NaCl, 1 mM ZnCl2.
Note: Also active in Low & Medium Buffers.
Assay Conditions: 30 mM sodium acetate (pH 5.0 at 22°C), 50 mM NaCl, 1 mM ZnCl2, 0.5 mg/ml denatured calf thymus DNA and 5% glycerol. Incubation is at 37°C for 10 min in a reaction volume of 1 ml.
Quality Control: All preparations are assayed for contaminating double-stranded DNase activity.
References:
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Ardelt, W., and Laskowski, M., Sr. (1971) Biochem. Biophys. Res. Commun. 44, No. 5, 1205-1211.
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Berk, A.J., and Sharp, P.A. (1977) Cell 12, 721-732.
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Berk, A.J., and Sharp, P.A. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 1274-1278.
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Goodman, H.M. and McDonald, R.J. (1979) Methods Enzymol. 68, 75-90.