Gene Expression Analysis in Mammalian Oocytes and Embryos by Quantitative Real-Time RT-PCR

Kyeoung-Hwa Kim; Su-Yeon Lee; Kyung-Ah Lee

doi:10.1007/978-1-4939-6988-3_4

Gene Expression Analysis in Mammalian Oocytes and Embryos by Quantitative Real-Time RT-PCR

Methods Mol Biol. 2017:1605:45-62. doi: 10.1007/978-1-4939-6988-3_4.

Authors

Kyeoung-Hwa Kim¹, Su-Yeon Lee¹, Kyung-Ah Lee²

Affiliations

¹ Department of Biomedical Science, Institute of Reproductive Medicine, College of Life Science, CHA University, Pangyo-Ro 335, Pan-Gyo, Korea.
² Department of Biomedical Science, Institute of Reproductive Medicine, College of Life Science, CHA University, Pangyo-Ro 335, Pan-Gyo, Korea. leeka@cha.ac.kr.

PMID: 28456957
DOI: 10.1007/978-1-4939-6988-3_4

Abstract

This chapter describes methods for preparing oocytes and embryos to analyze their gene expression at specific developmental stages. We illustrate how to collect germinal vesicles (GVs) and mature metaphase II (MII) stage oocytes, as well as how to collect embryos at specific developmental stages from the pronucleus (PN) to the blastocyst stage from female mice. We also describe how to prepare mRNAs from these precious cells to analyze the expression of the target genes. The materials and methods in this chapter are used mainly for mouse oocytes and embryos, but with subtle modifications, they may be applicable for most mammalian species.

Keywords: Gene expression; Oocytes; Pronuclear stage embryos; Zygotes; qRT-PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blastocyst / cytology*
Embryonic Development
Female
Gene Expression Profiling / methods*
Gene Expression Regulation, Developmental
Mice
Oocytes / cytology*
Oocytes / growth & development
RNA, Messenger / genetics*
Real-Time Polymerase Chain Reaction

Substances

RNA, Messenger