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Nucleic Acids Research, 2001, Vol. 29, No. 21 e106
© 2001 Oxford University Press

Bisulfite genomic sequencing of microdissected cells

Antoine Kerjean1,2,*, Annick Vieillefond3, Nicolas Thiounn4, Mathilde Sibony5, Marc Jeanpierre1 and Pierre Jouannet2

1Institut Cochin de Génétique Moléculaire, 2Laboratoire de Biologie de la Reproduction, 3Laboratoire d’Anatomie Pathologique and 4Clinique chirurgicale d’Urologie, Hôpital Cochin, 75014 Paris, France and 5Laboratoire d’Anatomie Pathologique, Hôpital Tenon, 75019 Paris, France

Mapping of methylation patterns in CpG islands has become an important tool for understanding tissue-specific gene expression in both normal and pathological situations. However, the inherent cellular heterogeneity of any given tissues can affect the outcome and interpretation of molecular studies. In order to analyse genomic DNA methylation on a pure cell population from tissue sample, we have developed a simple technique of single-cell microdissection from cryostat sections which can be combined with bisulfite-mediated sequencing of 5-methylcytosine. We report here our results on the methylation status of the androgen receptor gene studied by bisulfite genomic sequencing on purified cells isolated from human testis.

* To whom correspondence should be addressed at: INSERM U257, Institut Cochin de Génétique Moléculaire, 24 rue du Faubourg Saint-Jacques, 75014 Paris, France. Tel: +33 1 44 41 24 56; Fax: +33 1 44 41 24 62; Email: kerjean{at}cochin.inserm.fr


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