Mung Bean Nuclease
Mung Bean Nuclease
- Removal of hairpin loops during cDNA synthesis.1
- High-resolution mapping of the termini and exon structures of RNA transcripts (commonly termed Berk-Sharp or S1 Mapping) using either internal-labeled or end-labeled probes.2-5
- Restriction-site modification or removal by digestion of single-stranded protruding ends.2
- Cleavage of single-basepair mismatches, as a replacement for CEL 1 Nuclease in TILLING.6
- Unidirectional deletion of large DNA (in combination with Exonuclease III) to generate ordered deletions for sequencing.7
Mung Bean Nuclease is a single-strand-specific nuclease purified from sprouts of the mung bean Vigna radiata. Because Mung Bean Nuclease has higher specificity for ssDNA and RNA than S1 Nuclease, it is the enzyme of choice for most applications requiring a single-strand-specific nuclease. Unlike S1 Nuclease, Mung Bean Nuclease will not cleave the intact strand of nicked duplex DNA.
Unit Definition: One unit of Mung Bean Nuclease converts 1 µg of heat-denatured calf thymus DNA into an acid-soluble form in 1 minute at 37°C under standard assay conditions.
Storage Buffer: 50% glycerol containing 10 mM Tris-HCl (pH 7.5), 50 mM NaCl, and 0.01% Triton® X-100.
Mung Bean Nuclease 10X Reaction Buffer: 300 mM sodium acetate (pH 4.6), 500 mM NaCl, 10 mM zinc acetate, and 0.1% Triton® X-100.
Quality Control: Mung Bean Nuclease is tested in several functional assays, including precise deletion of ssDNA from cleaved restriction sites and nonrandom cleavage of partially denatured duplex DNA. The enzyme is also tested to ensure that the level of double-stranded nuclease activity is less than 0.05% of the single-stranded nuclease activity, and it is free of phosphatase activity.
- Gubler, U. (1987) Meth. Enzymol. 152, 330.
- Sambrook, J. et al. (1989) in: Molecular Cloning: A Laboratory Manual (2nd ed.), Cold Spring Harbor Laboratory Press, New York.
- Green, M.R. and Roeder, R.G. (1980) Cell 22, 231.
- Mathis, D.J. et al. (1981) Proc. Natl. Acad. Sci. USA 78, 7383.
- Murray, M.G. et al. (1986) Anal. Biochem. 158, 165.
- Till, BJ. et al. (2004) Nucleic Acids Res. 32, 2632.
- Henikoff, S. (1984) Gene 28, 351.