title: GSM9264345: Oocytes isolated from encapsulated follicles cultured in Blebbistatin - replicate 1; Mus musculus; RNA-Seq
library_name: GSM9264345
library_construction_protocol: We identified encapsulated follicles in the different culture conditions that grew beyond 180 µm and removed the alginate from them. Immediately after retrieval, the follicles were mechanically separated into oocytes and granulosa cells (GCs) within 5–10 minutes. While 1-2 oocytes were pooled together, a few hundred GCs were pooled into one sample set respectively. This was repeated individually for the different conditions to maintain similar levels of harvested RNA in each case. Each set of samples was promptly transferred into lysis buffer from the RNeasy Plus Micro Kit (Qiagen) and stored at –80 °C until RNA isolation. To minimize technical variability, RNA extraction was performed simultaneously for all biological replicates after the completion of all experiments.  Total RNA was extracted using the RNeasy Plus Micro Kit (Qiagen), following the manufacturer's instructions, and eluted in RNase-free water before being stored at –80 °C. Sequence-ready libraries were prepared following the manufacturer's instructions. Each sample utilized 8 µL of total RNA for cDNA synthesis, with cDNA amplification performed over 15 PCR cycles. Libraries were indexed using the NEBNext Multiplex Oligos for Illumina (NEB, E6440L)
identifiers.namespace: GEO
identifiers.id: GSM9264345_r1