GSM9039637: Starting pulmonary dissociation, replicate 1; Rattus norvegicus; RNA-Seq
Single-cell suspensions were prepared from dissociated organoids, bronchoalveolar lavage (BAL), and pulmonary dissociation (PD) samples using customized enzymatic or mechanical protocols, followed by filtration to achieve a single-cell suspension. Cells were resuspended in 0.1% BSA in PBS at 1 million cells/mL and processed for single-cell RNA-seq using the Chromium Next GEM Single Cell 3' v3.1 platform (10x Genomics, Pleasanton, CA), following the manufacturer's protocol. Cell lysis and mRNA capture occurred within Gel Beads-in-emulsion (GEMs) using barcoded oligo(dT) primers. See manuscript for more detail. Libraries were prepared using the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10x Genomics, Pleasanton, CA), following the manufacturer's protocol. Barcoded cDNA was amplified by PCR, purified with SPRIselect beads, and prepared for sequencing on an Illumina platform. See manuscript for more detail.