A crude nuclease preparation suitable for use in DNA reassociation experiments

Sutton, W.D.

doi:10.1016/0005-2787(71)90709-x

Cited by 413 publications

(85 citation statements)

References 28 publications

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“…Nuclease S, action was found to depend upon ionic strength as reported by Sutton (1971). In our case, the rate of DNA hydrolysis in the presence of 0.03 M-NaCl was 94 O 0 of that in the absence of NaCI, and the inhibitory effect of NaCl on DNA hydrolysis increased with the NaCl concentration.…”

Section: Dna Digestion By Nuclease Ssupporting

confidence: 83%

Classification of Micrococci on the Basis of Deoxyribonucleic Acid Homology

Ogasawara-Fujita¹,

Sakaguchi²

1976

Journal of General Microbiology

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S U M M A R YThe DNA homology relationships of 25 micrococci (15 strains of Micrococcus, eight strains of Sarcina and two strains of Staphylococcus) were studied by the deoxyribonucleic acid hybridization method using nuclease S,, an endonuclease specific for single-stranded DNA molecules. Nineteen of the strains were classified into three groups. De Ley et al., 1973). The DNA hybridization method using nitrocellulose membranes is common today, but this method has the disadvantage that DNA fixed on nitrocellulose membranes is released during incubation at high temperatures (Okanishi & Gregory, I 9 7 0 ) . Recently, nuclease S, isolated from Aspergillus oryzae, which is specific for single-stranded nucleic acid (Ando, I 966), was successfully used in improving the nitrocellulose-membrane DNA hybridization method. This endonuclease digests only the single-stranded DNA portion that has not hybridized. Crosa, Brenner & Falkow (I 973) studied the nucleotide sequence relatedness between homologous and heterologous chromosomal and plasmid DNA by the DNA hybridization method with nuclease S, and showed that this method permitted an accurate, reproducible and rapid determination of the extent of polynucleotide sequence homology. In this study, we examined the DNA homology relationships, by DNA hybridization followed by nuclease S, digestion, among species of the genus Micrococcus, including aerobic Sarcina strains which had been proved to belong to the genus Micrococcus by Kocur & Martinec (1965) and Hubalek (1969). 7-2

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Section: Dna Digestion By Nuclease Ssupporting

confidence: 83%

Classification of Micrococci on the Basis of Deoxyribonucleic Acid Homology

Ogasawara-Fujita¹,

Sakaguchi²

1976

Journal of General Microbiology

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“…an equal volume of 005 M sodium acetate buffer pH 43 containing 02 mrvi Zn++ and 55 mat mercaptoethanol was added to bring the pH to 43. Enough S nuclease, purified by the method of Sutton (1971), was added to be able to digest all the DNA, if it were single stranded, during a 60 mm. incubation at 37°C.…”

Section: Materials and Methods (I) Plant Genotypesmentioning

confidence: 99%

Repeated sequence DNA comparisons between Triticum and Aegilops species

1979

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SUMMARYThe families of repeated sequences in the genomes of a range of Triticum and Aegilops species have been compared. All the genomes are very similar. However, using a DNA probe from Aegilops speltoides that contains the most highly repeated sequences, diploid Aegilops species could be distinguished from diploid Triticum species. Different Ae'gilops species' DNAs also hybridise to differing extents with this probe. The results are consistent with the hypothesis that speciation is accompanied by quantitative changes in the repeated sequence complements of genomes.Most if not all of the families of repeated sequences in hexaploid wheat can be detected in Aegilops speltoia'es (related to the B genome) and in Aegilops squarrosa (related to the D genome). However, some families of repeated sequences of hexaploid wheat were not found in Triticum monococcum (related to the A genome). Some of the most highly repeated sequences of hexaploid wheat are preferentially concentrated in the B genome. These sequences are useful as probes for distinguishing the three diploid genomes of hexaploid wheat.

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“…(vi) S1 nuclease digestion S1 nuclease was prepared from "Takadiastase" (Koch Light Ltd) by the method of Sutton (1971) and stored in 50 per cent glycerol at -20°C.…”

Section: Materials and Methods (I) Isolation Of 3h-labelled And Unlabmentioning

confidence: 99%