title: GSM8991353: MOS_mutant_M1_replicate_8; Mus musculus; RNA-Seq
library_construction_protocol: Ovaries were dissected, and germinal vesicle (GV) oocytes were isolated in minimal essential medium supplemented with 2.5μM milrinone to prevent meiotic resumption.  To induce synchronous meiotic progression, oocytes were then cultured in milrinone-free Chatot, Ziomek, and Bavister (CZB) medium in a humidified incubator at 37°C with 5% CO₂. For chemical activation, wildtype oocytes were matured in CZB for 15-16 hours after germinal vesicle breakdown, then incubated for 2 hours in calcium-free CZB containing 10 mM strontium chloride. Following activation, the eggs were washed and maintained in CZB. scRNA-seq libraries were generated using SMART-Seq2 protocol (Trombetta et al., 2014). Briefly, cDNA was reversed transcribed from single cells using SuperScript III (Thermo Fisher Scientific) and whole transcriptome amplification (WTA) was performed. WTA products were purified using the Agencourt AMPure XP beads (Beckman Coulter, IN, USA) and used to prepare paired-end libraries with Nextera XT  (Illumina, CA, USA).