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Science 5 September 1997:
Vol. 277. no. 5331, pp. 1518 - 1523
DOI: 10.1126/science.277.5331.1518
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Reports

Dynamic Molecular Combing: Stretching the Whole Human Genome for High-Resolution Studies

Xavier Michalet, * Rosemary Ekong, * Françoise Fougerousse, Sophie Rousseaux, dagger Catherine Schurra, Nick Hornigold, Marjon van Slegtenhorst, Jonathan Wolfe, Sue Povey, Jacques S. Beckmann, Aaron Bensimon ddagger

DNA in amounts representative of hundreds of eukaryotic genomes was extended on silanized surfaces by dynamic molecular combing. The precise measurement of hybridized DNA probes was achieved directly without requiring normalization. This approach was validated with the high-resolution mapping of cosmid contigs on a yeast artificial chromosome (YAC) within yeast genomic DNA. It was extended to human genomic DNA for precise measurements ranging from 7 to 150 kilobases, of gaps within a contig, and of microdeletions in the tuberous sclerosis 2 gene on patients' DNA. The simplicity, reproducibility, and precision of this approach makes it a powerful tool for a variety of genomic studies.

X. Michalet, C. Schurra, A. Bensimon, Laboratoire de Biophysique de l'ADN, Département des Biotechnologies, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.
R. Ekong, S. Rousseaux, S. Povey, MRC Human Biochemical Genetics Unit, University College London, 4 Stephenson Way, London NW1 2HE, UK.
F. Fougerousse and J. S. Beckmann, URA 1922, Généthon, 1 rue de l'Internationale, Boîte Postale 60, 91002 Evry, France.
N. Hornigold and J. Wolfe, The Galton Laboratory, Department of Biology, University College London, 4 Stephenson Way, London NW1 2HE, UK.
M. van Slegtenhorst, Department of Clinical Genetics, Erasmus University, Dr. Molewaterplein 50, NL-3015, GE Rotterdam, Netherlands.
*   These authors contributed equally to this work.

dagger    Present address: INSERM U309, Institut Albert Bonniot, 39706 La Tronche Cedex, France.

ddagger    To whom correspondence should be addressed. E-mail: abensim{at}pasteur.fr

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