DESCRIPTION
Mung Bean Nuclease degrades single-stranded nucleic acids to yield 5'-phosphoryl mono- or oligonucleotides. Double-stranded DNA, RNA and DNA:RNA hybrids are relatively resistant to the enzyme. If very large amount of enzyme are used it also degrades double-stranded DNA from both ends. Unlike S1 nuclease, this enzyme will not cleave the DNA strand opposite a nick in a duplex.
SOURCE
Mung bean sprouts.
APPLICATIONS
¡Ü Removal of 3' and 5' extensions from DNA or RNA
termini.
¡Ü Transcript mapping.
¡Ü Opening up hairpin loops formed during synthesis of
double-stranded cDNA.
¡Ü Excision of gene coding sequences from genomic
DNA.
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UNIT DEFINITION
One unit is defined as the amount of enzyme required to produce 1¥ìg of acid-soluble nucleotides 1 minutes at 37¡É.
STANDARD UNIT ASSAY CONDITIONS
30mM sodium acetate (pH4.6), 50mM NaCl, 1mM ZnCl2, 0.5mg/ml denatured calf thymus DNA and 5%(v/v) glycerol.
CONCENTRATION: 50-100 units/ul.
10X REACTION BUFFER
300mM sodium acetate(pH5.0), 500mM NaCl, 10mM ZnCl2.
STORAGE CONDITIONS
10mM sodium acetate (pH5.0), 0.1mM zinc acetate, 1mM cysteine, 0.001% Triton X-100, and 50% glycerol. Store at -20¡É.
REFERENCES
1. Kowalski, D. et al. (1976) Biochemistry 15, 4457.
2. Green, M.R. and Roeder, R.G. (1980) Cell 22, 231.
3. Gubler, U. (1987) Methods Enzymol. 152, 330.
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