GSM5348650: OMIIO_2; Mus musculus; RNA-Seq
Ovaries were separated and transferred into M2 medium (Sigma). For oocyte and GCs at growing stage (GO and GCs), the follicles with a diameter of 200-250 μm were isolated using insulin syringe under stereoscope. Full-grown oocytes and GCs (FGO and FGCs) were obtained 48 h after pregnant mare serum gonadotropin (PMSG) priming. For oocyte and GCs at MI stage (MIO and MIGCs), C57BL/6 female mice were intraperitoneally injected with 10 units PMSG, and 48 h later injected with 10 units human chorionic gonadotropin (hCG). After 9 h, the mice were sacrificed by cervical dislocation and the ovaries were separated. The pre-ovulatory follicles were punctured using a 1 mL disposable syringe under stereoscope, and COCs were transferred into new hyaluronidase droplets with the mouth pipette for 5 minutes of digestion. For oocyte and GCs at MII stage (MIIO and MIIGCs), the COCs were isolated from oviduct after 13 h and cumulus masses were separated in hyaluronidase medium. Oocytes and GCs from young (6 weeks old ) and aged (10 months old) mice at four stages were collected and processed for smart RNA-seq. RNA-seq libraries were constructed as previously described (Liu et al., 2021). Discover-scTM WTA Kit V2 (Vazyme, N711) was used to obtain cDNA from single oocyte and GCs. The TruePrepTM DNA Library Prep Kit V2 for Illumina (Vazyme, TD503) was used for the purpose of constructing libraries. An Illumina HiSeq X platform (Shanghai, China) performs libraries sequencing. High-quality reads were aligned to Mus musculus UCSC mm9 references and FPKM value of each gene was calculated.