Analysis of DNA methylation patterns in single blastocysts by Pyrosequencing(®).

John Huntriss, Kathryn Woodfine, Joanna E. Huddleston, Adele Murrell, Helen M. Picton

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Citations (Scopus)

Abstract

Extensive epigenetic reprogramming occurs during mammalian gametogenesis and preimplantation development. DNA methylation patterns that are laid down during these stages are essential for subsequent normal foetal development. The requirement for more precise assessment of the epigenetic programming of in vitro-derived human preimplantation embryo has become of paramount importance following the identification of epigenetic diseases that are associated with assisted reproduction and/or infertility. Such techniques are also useful and applicable to experimental reproductive biology. In order to expand our knowledge of epigenetic marks, including DNA methylation, during mammalian reproduction and early development, it is necessary to test new and sufficiently sensitive protocols. There are, however, unique challenges to obtain DNA methylation data from the small cell numbers that are present in the preimplantation embryo. In this protocol, we describe the successful application of Pyrosequencing(®) to yield quantitative DNA methylation data over several CpG sites at differentially methylated regions (DMRs) at imprinted loci in single blastocysts, in this case, human blastocysts. Future developments of the protocol will allow DNA methylation analysis of a more extensive panel of genes for each embryo and at the same time, since the protocol allows for the extraction of mRNA from the embryo, the comparison between DNA methylation and gene expression
Original languageEnglish
Title of host publicationPyrosequencing
Subtitle of host publicationMethods and Protocols - Part III
EditorsU. Lehmann, J. Tost
Place of PublicationNew York, U. S. A.
PublisherSpringer
Pages259-270
ISBN (Print)9781493927142
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology
Volume1315

Fingerprint

DNA methylation
blastocyst
epigenetics
embryo (animal)
assisted reproductive technologies
fetal development
gametogenesis
early development
Biological Sciences
gene expression
loci
genes
testing
cells

Cite this

Huntriss, J., Woodfine, K., Huddleston, J. E., Murrell, A., & Picton, H. M. (2015). Analysis of DNA methylation patterns in single blastocysts by Pyrosequencing(®). In U. Lehmann, & J. Tost (Eds.), Pyrosequencing: Methods and Protocols - Part III (pp. 259-270). (Methods in Molecular Biology; Vol. 1315). New York, U. S. A.: Springer. https://doi.org/10.1007/978-1-4939-2715-9_19

Analysis of DNA methylation patterns in single blastocysts by Pyrosequencing(®). / Huntriss, John; Woodfine, Kathryn; Huddleston, Joanna E.; Murrell, Adele; Picton, Helen M.

Pyrosequencing: Methods and Protocols - Part III. ed. / U. Lehmann; J. Tost. New York, U. S. A. : Springer, 2015. p. 259-270 (Methods in Molecular Biology; Vol. 1315).

Research output: Chapter in Book/Report/Conference proceedingChapter

Huntriss, J, Woodfine, K, Huddleston, JE, Murrell, A & Picton, HM 2015, Analysis of DNA methylation patterns in single blastocysts by Pyrosequencing(®). in U Lehmann & J Tost (eds), Pyrosequencing: Methods and Protocols - Part III. Methods in Molecular Biology, vol. 1315, Springer, New York, U. S. A., pp. 259-270. https://doi.org/10.1007/978-1-4939-2715-9_19
Huntriss J, Woodfine K, Huddleston JE, Murrell A, Picton HM. Analysis of DNA methylation patterns in single blastocysts by Pyrosequencing(®). In Lehmann U, Tost J, editors, Pyrosequencing: Methods and Protocols - Part III. New York, U. S. A.: Springer. 2015. p. 259-270. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-2715-9_19
Huntriss, John ; Woodfine, Kathryn ; Huddleston, Joanna E. ; Murrell, Adele ; Picton, Helen M. / Analysis of DNA methylation patterns in single blastocysts by Pyrosequencing(®). Pyrosequencing: Methods and Protocols - Part III. editor / U. Lehmann ; J. Tost. New York, U. S. A. : Springer, 2015. pp. 259-270 (Methods in Molecular Biology).
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