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.
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. |