GSM9428634: Zhedong White goose thymus, embryonic day 25 (E25), scRNA-seq; Anser cygnoides; RNA-Seq
Thymus tissues were minced on ice, washed with 1× PBS, and dissociated into single cells using a dissociation solution at 37 °C for 20 min with gentle shaking. Digestion was terminated by adding PBS containing 10% fetal bovine serum (FBS). Cell suspensions were filtered through a 40-μm cell strainer, centrifuged at 300 × g for 5 min at 4 °C, and resuspended in PBS supplemented with 0.04% BSA. Residual red blood cells were removed using 1× red blood cell lysis buffer. Cell viability was assessed by trypan blue exclusion, and only samples with viability greater than 85% were used for downstream processing. Single-cell RNA-seq libraries were prepared using the 10× Genomics Chromium platform with the Chromium Single Cell 3′ Library Kit (v3), following the manufacturer's instructions. Single-cell suspensions were adjusted to a concentration of 400–1,200 cells/μL prior to library construction. Libraries were sequenced on an Illumina NovaSeq 6000 system in paired-end 150 bp mode.