Mung Bean Nuclease

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Last modified: 2008-07-15 14:57:15

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 single-stranded DNA or 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. Because of these characteristics, it is preferable to S1 Nuclease for many applications.

Unit Definition:

One unit converts 1µg of heat-denatured calf thymus DNA to an acid-soluble form in 1 minute at 37°C in 30mM sodium acetate, pH 4.6, 50mM NaCl, 1mM zinc acetate, and 0.01% Triton® X-100.

Storage Buffer:

50% glycerol containing 10mM Tris-HCl, pH 7.5, 50mM NaCl, and 0.01% Triton® X-100.

Mung Bean Nuclease 10X Reaction Buffer: 300 mM sodium acetate, pH4.6, 500mM sodium chloride, 10mM zinc acetate, and 0.1% Triton® X-100.

Quality Control:

Mung Bean Nuclease is tested in several functional assays, including precise deletion of single-stranded DNA from cleaved restriction sites and non-random 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.

Last modified: 2008-05-14 16:46:46

Applications

• Removal of hairpin structures during cDNA synthesis
• High resolution mapping of the termini and exon structures of RNA transcripts (commonly termed Berk-Sharp or S1-Mapping) using either internally-labelled
• Restriction site modification or removal by digestion of single-stranded protruding ends
• Unidirectional deletion of large DNA (in combination with Exo III) to generate ordered deletions for sequencing (i.e., an alternative to nuclease BAL-31 deletions)

Last modified: 2008-05-14 16:46:46

Protocols for: Mung Bean Nuclease

(catalogue number M8202K / M8205K)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46

References

1. Gubler, U. (1987) Meth. Enzymol. 152, 330.
2. Sambrook, J. et al. (1989) in: Molecular Cloning: A Laboratory Manual (2nd ed.), Cold Spring Harbor Laboratory Press, New York.
3. Green, M.R. and Roeder, R.G. (1980) Cell 22, 231.
4. Mathis, D.J. et al. (1981) Proc. Natl. Acad. Sci. USA 78, 7383.
5. Murray, M.G. et al. (1986) Anal. Biochem. 158, 165.
6. Till, BJ. et al. (2004) Nucleic Acids Res. 32, 2632.
7. Henikoff, S. (1984) Gene 28, 351.

Last modified: 2008-05-14 16:46:46