Bovine oocytes matured with 0.1% ethanol
Ethanol is a potential risk factor affecting oocyte quality in humans. This data is RNAseq of metaphase 2 stage oocytes matured with 0.1% ethanol. Ovaries were collected at a slaughterhouse and transported to the laboratory within 4 hours. Oocyte-cumulus cells complexes (COCs) were collected from bovine antral follicles (3-5 mm in diameter) of Japanese Black Cows and cultured in TCM-199 medium containing 10ng/ml EGF, 10% FCS and with or without 0.1% ethanol. At the end of culture period (21hrs), oocytes were denuded from surrounding cumulus cells were washed in PBS containing 0.1%PVA. The RNA extracted from the 50 oocytes were subjected to RNA seq. RNA extraction was conducted using RNAqueous Kit(Invitrogen). Three samples were prepared from differential ovarian batches. 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 manufacturers instructions, and the sequence data was converted to Fastq using bcl2fastq2 v2.20.0.422.