GSM8842800: 10x single-cell RNA-seq bat small intestinal organoids; Rousettus aegyptiacus; RNA-Seq
Single cell suspension were obtained by enzymatic digestion of tissue (collagenase from Clostridium histolyticum and Dispase) or organoids (TrypLE Express) followed by AnnexinV magnetic bead-based selection of viable cells (STEMCELL technologies). Cells were resuspended in DMEM/F12 and submitted to an on-campus facility for 10x library preparation. For bulk RNA sequencing, total RNA was extracted using a magnetic bead-based semi-automatic purification system (Kingfisher). Single cell RNA-sequencing Libraries were generated according to the 10x Chromium Single Cell 3' Reagent Kit v3 and v4 kit manual instructions. Pooled 3'-end bulk RNA-sequencing libraries were prepared using an in-house protocol that adheres to the PLATE-seq (PMID: 28740083; modified to include UMIs and optimized steps for library generation) and BRB-seq (PMID: 30999927; for bioinformatic tools) methodologies (see methods of associated publication).