Mung Bean Nuclease

Mung Bean Nuclease belongs to a class of enzymes that demonstrate a preference for single-stranded nucleic acids, lack sugar specificity and hydrolyze single-stranded substrates to products with 5пїЅ-phosphoryl and 3пїЅ-hydroxyl termini. Mung Bean Nuclease catalyzes the degradation of single-stranded DNA and RNA endonucleolytically to yield 5'-phosphoryl terminated products. While the nuclease prefers ssDNA over dsDNA by 30,000-fold, at very high concentrations, such as 14u/ug of DNA, the enzyme degrades double-stranded DNA preferentially from both ends (1,2,3).\n\nUnit Definition: One unit is defined as the amount of enzyme required to produce 1ug of acid-soluble nucleotides per minute at 37 degrees C in 30mM sodium acetate, pH 5.0, 50mM NaCl, 1mM ZnCl2, 0.5mg/ml denatured calf thymus DNA, 5% glycerol.\n\nAdditional Information\nMolecular Weight: 39kD\nRequirement: Zn2+.\nInhibitors: Mung Bean Nuclease is inhibited by high salt concentrations (80-90% inhibition in 200-400mM NaCl). Use with 0.001% Triton(R) X-100 when using very low concentrations (< 50u/ml), because under such conditions Mung Bean Nuclease may adhere to surfaces and is rather unstable (7).\n\nQuality Control:\nContaminant Endonuclease/Nickase Activity: To confirm the absence of contaminating endonuclease/nickase activity, 1ug of lambda DNA/ Hind III markers is incubated with Mung Bean Nuclease for 10 minutes at RT. The markers are then separated by electrophoresis on a 1% agarose gel and stained with ethidium bromide. Markers that have been incubated with (same/more than) 60u of enzyme will remain as intact bands with a minimal amount of smearing.\n\nApplications:\nSuitable for use in Transcript Mapping, Production of blunt ends for cloning, Flushing staggered ends and Separation of cDNA strands after synthesis with reverse transcriptase and DNA polymerase I.\n\nReaction Buffer:\nM9218A: Mung Bean Nuclease, Reaction Buffer (10X), 1x1ml\nForm: Supplied as a liquid in 0.3Msodium acetate, pH 5.0, 0.5M sodium chloride, 10mM ZnCl2.\nRecommended Dilution: 1:10\n\nStorage and Stability:\nAliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

M9218

Size

2000U

Form

Supplied as a liquid in 0.01M Tris-HCl, pH 7.5, 0.05M sodium chloride, 50% glycerol, 0.01% Triton X-100.

References

1. Ardelt, W., Laskowski, M. Sr.: Mung bean nuclease I. IV. An improved method of preparation. Biochem. Biophys. Res. Comm. 44: 1205 (1971). 2. Kroeker, W.D., Kowalski, D., Laskowski, M. Sr.: Biochemistry15: 4463 (1976). 3. Kroeker, W., Spurgeon, S.: Mung bean nuclease (1985). 4. Mathis, D.J., et al.: Specific in vitroinitiation of transcription on the adenovirus type 2 early and late EII transcription units. PNAS USA 78: 7383 (1981). 5. Green, M.R., Roeder, R.G.: Definition of a novel promoter for the major adenovirus-associated virus mRNA. Cell 22: 231 (1980). 6. Gubler, U.: Second-strand cDNA synthesis: mRNA fragments as primers. Meth. Enzymol. 152: 330 (1987). 7. Perbal, B.: A Practical Guide to Molecular Cloning, 2nd ed., John Wiley and Sons, New York (1988).