DESCRIPTION
S1 Nuclease degrades single- stranded DNA or RNA to yield 5'-phosphate mono- or oligonucleotides. Double-stranded DNA, double-stranded RNA, and DNA:RNA hybrids are relatively resistant to the degradation. Double-stranded nucleic acid are digested completely by S1 nuclease in the presence of very large amount of enzyme.
SOURCE
Aspergillus oryzae.
APPLICATIONS
¡Ü S1 mapping of RNA transcripts.
¡Ü Removing single-stranded overhangs from DNA fragments to produce blunt ends.
¡Ü Opening up the hairpin loops generated during synthesis of double-stranded cDNA.
UNIT DEFINITION
One unit is defined as the amount of enzyme that produces 1¥ìg of acid-soluble material per minute at 37¡É.
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STANDARD UNIT ASSAY CONDITIONS
30mM Sodium acetate(pH4.6), 50mM NaCl, 1mM zinc chloride, 0.5mg/ml denatured calf thymus DNA and 5% glycerol.
CONCENTRATION: 20-100 units/¥ìl.
10X REACTION BUFFER
500mM Sodium acetate (pH4.5), 2800mM NaCl, 45mM zinc sulphate.
STORAGE CONDITIONS
20mM Tris-HCl (pH7.5), 50mM NaCl, 0.1mM zinc sulphate, and 50% glycerol. Store at -20¡É.
REFERENCES
1. Berk, A. J. and Sharp, P. A. (1978) Proc. Natl. Acad. Sci. USA 75, 1274.
2. Vogt, V. M. (1973) Eur. J. Biochem. 33, 192.
3. Roberts, T. M. et al. (1979) Proc. Natl. Acad. Sci. USA 76, 760.
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