|
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.
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.
|