Gene expression analysis in mammalian oocytes and embryos by quantitative real-time RT-PCR

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.