Antonie van Leeuwenhoek

, Volume 36, Issue 1, pp 461–474

Evaluation of membrane filter methods for DNA-DNA hybridization

Authors

  • J. De Ley
    • Laboratory for Microbiology and Microbial Genetics, Faculty of SciencesState University
  • R. Tijtgat
    • Laboratory for Microbiology and Microbial Genetics, Faculty of SciencesState University
Article

DOI: 10.1007/BF02069048

Cite this article as:
De Ley, J. & Tijtgat, R. Antonie van Leeuwenhoek (1970) 36: 461. doi:10.1007/BF02069048

Abstract

Three membrane filter methods for DNA hybridization were compared. In all of them filter-fixed DNA is spontaneously released during incubation. The effect depends largely on the temperature: 40 to 85% of DNA is released at 70 to 80 C; at higher temperatures most of the DNA is released and heterologous DNA relatedness values are unreliable. Only the dimethylsulfoxide filter method (Legault-Démare et al., 1967) can be recommended for the entire temperature and % GC range.

The methods of Denhardt (1966) and of Warnaar and Cohen (1966) are not recommended for hybridizations of DNA containing more than 50% GC. Impure DNA yields too low DNA relatedness values.

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Copyright information

© Swets & Zeitlinger 1970