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Nucleic Acids Research, 2000, Vol. 28, No. 10 E48-e48
© 2000 Oxford University Press

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

Jean-François Brugère, Emmanuel Cornillot*, Guy Méténier and Christian P. Vivarès

Equipe de Parasitologie Moléculaire et Cellulaire, UPRES A CNRS 6023, Université Blaise Pascal, 63177 Aubière cedex, France

A simple method for complete genome radiolabelling is described, involving long-wave UV exposure of agarose-embedded chromosomal DNA and [{alpha}-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.

* To whom correspondence should be addressed. Tel: +33 4 73 40 74 71; Fax: +33 4 73 40 74 55; Email: emmanuel.cornillot@lbp.univ-bpclermont.fr


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