Methylation of Specific Regions: Bisulfite-Sequencing at the Single Oocyte or 2-Cell Embryo Level
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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 wordsOocyte DNA methylation PCR at the single cell level
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