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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The senior author (J. D. L.) is indebted to the Fonds voor Kollektief Fundamenteel Onderzoek for research and personnel grants, and to Dr N. Walker, Rothamsted Experimental Station, UK, for kindly correcting language and style errors in the typescript.
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De Ley, J., Tijtgat, R. Evaluation of membrane filter methods for DNA-DNA hybridization. Antonie van Leeuwenhoek 36, 461–474 (1970). https://doi.org/10.1007/BF02069048
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DOI: https://doi.org/10.1007/BF02069048