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Oxford University Press, Nucleic Acids Research, 10(28), p. 48e-48, 2000

DOI: 10.1093/nar/28.10.e48

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In-gel DNA radiolabelling and two-dimensional pulsed field gel electrophoresis procedures suitable for fingerprinting and mapping small eukaryotic genomes

Journal article published in 2000 by Emmanuel Cornillot, Guy Méténier, Christian P. Vivarès, Jean-François Brugère ORCID
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

A simple method for complete genome radiolabelling is described, involving long-wave UV exposure of agarose-embedded chromosomal DNA and [α-32P]dCTP incorporation mediated by the Klenow fragment. Experiments on the budding yeast genome show that the labelling procedure can be coupled with two new two-dimensional pulsed field gel electrophoresis (2D-PFGE) protocols of genome analysis: (i) the KARD (karyotype and restriction display)-PFGE which provides a complete view of the fragments resulting from a single restriction of the whole genome and (ii) the DDIC (double digestion of isolated chromosome)-PFGE which is the eukaryotic counterpart of complete/complete 2D-PFGE in bacterial genomics.