title: GSM4452395: DOs3_RNA-seq; Ovis aries; RNA-Seq
library_construction_protocol: The follicular oocytes of 2~8 mm were extracted from the washed ovaries, after settlement for a short time, the supernatant was removed, and cell pellet was resuspended with washing buffer for subsequent selection. Under stereoscopic microscope, the oocytes were divided into two classes, one is denuded oocyte (GV stage), other oocytes surrounded by granulosa cells (cumulus oocyte complexes, COCs)  were treated with hyaluronidase to remove cumulus cells, and cultured in vitro with maturation medium in a humidified incubator at 38.5 ℃ with 5% CO2 for 23-25 hours. The MⅡ stage oocytes were harvested with first polar body extrusion. The sequence samples were prepared with 30-35 oocytes as one biological repeat. With high quality of RNA, an amount of 3 µg RNA each sample was used as input to prepare sequence. After removement of ribosomal RNA, the sequence libraries were generated with NEB Next® Ultra™ Directional RNA Library Prep Kit (NEB, USA) according to manufacturer's recommendations, the quality of which was assessed with Agilent Bioanalyzer 2100 system. After samples clustering with TruSeq PE Cluster Kit v3-cBot-HS (Illumia) according to the manufacturer's instructions, sequence was conducted on Illumina Hiseq 2000 platform with 100 bp paired-end strategy.
attributes.tag: GEO Accession
attributes.value: GSM4452395