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Methylation of Specific Regions: Bisulfite-Sequencing at the Single Oocyte or 2-Cell Embryo Level

  • Annick Lefèvre
  • Thierry BlachèreEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1222)

Abstract

To question the possible implication of an alteration of the DNA methylation of imprinted genes in normal development failure observed following fertilization in ART centers, it has been necessary to develop a reproducible and highly efficient method to perform analysis at the one cell level. We have thus developed a very efficient protocol for methylation studies on individual oocytes or cleavage-stage embryos. All the different steps were optimized, from DNA extraction, to limit DNA degradation and give a high success rate of bisulfite converted DNA, to amplification of the bisulfite modified DNA.

Key words

Oocyte DNA methylation PCR at the single cell level 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Institut Cellule Souche et CerveauINSERM U846BronFrance
  2. 2.Laboratoire de Biologie de la reproduction, Faculté de MédecineINSERM U846Lyon Cedex 08France

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