GSM8254745: Postnatal day 20 female oocyte, Classified as SN, Rep1; Mus musculus; RNA-Seq
The oocytes were recovered from the ovaries by follicle puncture and cumulus oocyte complexes were transferred to minimal essential medium (MEM) supplemented with 3mg/ml bovine serum albumin (Sigma Aldrich, St. Louis, MO) and milrinone (Sigma; 1 μg/ml) to prevent meiotic resumption. Surrounding cumulus cells were removed by gentle pipetting followed by several rinses in fresh medium. Oocytes were cultured for specified periods of time and maintained at 37°C in MEM/BSA plus milrinone under 5% CO2, 5%O2 and balanced N2. For RNA-seq, oocytes are denuded in Tyrode's solution (Sigma) and transferred to microcentrifuge tubes containing 3 μl of lysis buffer. Isolation of mRNA was conducted using oligo(dT)