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Nucleic Acids Research, 1978, Vol. 5, No. 11 4283-4304
© 1978

Articles

Isolation of discrete repetitive sequence classes from Xenopus DNA by high temperature reassociation

Barbara A. Braun, Kathleen E. Schanke and Dale E. Graham

Department of Biological Sciences, Purdue University West Lafayette, IN 47907, USA

Received August 22, 1978. Sequences that did or did not reassociate at 75°C (stable and unstable, respectively) were isolated from total repetitive Xenopus laevis DNA. Sequence complexities or frequencies were determined by self (minicot) or DNA excess (slave minicot) reassociations at 60°C. Stable sequences were five times shorter and four times more frequent than unstable sequences. Reassociations at 75°C or at 50°C were used to establish apparent sequence frequencies at these criteria. Interspersion curves at either 60°C or 75°C and low Cot reassociation of long fragments of total X. laevis DNA at either 60°C or 75°C, followed by S1 digestion and agarose chromatography, were used to determine genome arrangement of the stable and unstable classes of sequence. Reassociation at high temperature was found to permit the fractionation of repetitive sequences into two populations of differing characteristics.


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