A crude nuclease preparation suitable for use in DNA reassociation experiments

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Abstract

A crude preparation of nuclease S1 from Aspergillus oryzae is shown to have a strong preference for heat-denatured DNA as substrate, to be active over a range of ionic strengths, to degrade a variety of single-stranded DNA's to better than 97 % acid-soluble materials, and to be capable of removing the single-stranded regions from reassociated DNA molecules at a range of temperatures without extensively degrading the reassociated duplex regions. Nuclease S1 displays anomalous reaction kinetics at low substrate concentrations.

References (33)

  • P.M.B. Walker
  • B.J. McCarthy et al.

    J. Mol. Biol.

    (1964)
  • D. Gillespie et al.

    J. Mol. Biol.

    (1965)
  • A.P. Nygaard et al.

    Biochem. Biophys. Res. Commun.

    (1963)
  • T. Ando

    Biochim. Biophys. Acta

    (1966)
  • P.M.B. Walker et al.

    J. Mol. Biol.

    (1965)
  • J. Marmur

    J. Mol. Biol.

    (1961)
  • W.G. Flamm et al.

    J. Mol. Biol.

    (1969)
  • W.G. Flamm et al.

    J. Mol. Biol.

    (1969)
  • F.W. Studier

    J. Mol. Biol.

    (1965)
  • S.C. Sung et al.

    J. Biol. Chem.

    (1962)
  • J. Brahms et al.

    J. Mol. Biol.

    (1966)
  • I.R. Lehman

    J. Biol. Chem.

    (1960)
  • I.R. Lehman et al.

    J. Biol. Chem.

    (1964)
  • C.L. Schildkraut et al.

    J. Mol. Biol.

    (1961)
  • P.H. Johnson et al.

    J. Biol. Chem.

    (1970)
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