title: GSM9159779: Juvenile_XL_ovary, biol_ rep_2; Xenopus laevis; RNA-Seq
library_construction_protocol: Ovaries from juvenile Xenopus laevis (NF62–66) were chopped and enzymatically dissociated in a collagenase mix (L-15 with 3 mg/ml Collagenase I [Gibco 17018029], 2 mg/ml Collagenase IV [Gibco 17104019], 3 mg/ml Collagenase II [Sigma C6885], and 1 mM CaCl₂) for 40 min at room temperature with rotation. Cells were washed in PBS, treated with TrypLE Express (Gibco) for 10 min, washed again, re-suspended in cold PBS, and filtered through 70 μm and 40 μm strainers. The final cell suspension was prepared in PBS with 1% BSA for 10X Genomics loading. Libraries were prepared using the 10X Genomics Chromium GEM-X Single Cell 3′ Reagent Kit v4, following the manufacturer's protocol. Approximately 10,000 cells per sample were loaded onto the Chromium platform to generate Gel Beads-in-Emulsion (GEMs), followed by reverse transcription, cDNA amplification, and library preparation according to the 10X workflow.