mouse oocytes matured with 0.2% ethanol

Ethanol is a potential risk factor affecting oocyte quality in humans. This data is RNA-seq of metaphase II stage oocytes matured with 0.2% ethanol. Ovaries were collected from 8week mouse (ICR). Oocyte-cumulus cells complexes (COCs) were collected from antral follicles and cultured in a-MEM containing 10mM Taurine, 5% FCS and 10uAU/ml FSH (Kyoritsu, Tokyo, Japan) and with or without 0.2% ethanol. At the end of culture period (21hrs), oocytes were denuded from surrounding cumulus cells and washed in PBS containing 0.1%PVA. The RNA extracted from the 44 oocytes were subjected to RNA-seq. RNA extraction was conducted using RNAqueous Kit (Invitrogen). Three samples were prepared from ovaries of at least three ICR mice. The RNA quality and concentration were examined using a Bioanalyzer (Agilent technologies, Palo Alto, CA, USA). cDNA of the oocytes was produced from each RNA using NEBNext Single Cell/Low Input RNA Library Prep Kit. The quality and quantity were determined using the Agilent 2100 Bioanalyzer, followed by re-measurement using Kapa Library Quantification Kit Kapa Biosystems. Sequencing was conducted using NextSeq1000 Single read x 100 bp. Image analysis, base calling and quality filtering were performed using the RTA version 2.4.11 (Illumina) following the manufacturer' s instructions, and the sequence data was converted to Fastq using bcl2fastq2 v2.20.0.422.