Single-cell RNA-seq using Chromium Platform

The scRNA-seq libraries were prepared using Chromium Single Cell 3 Reagent v3 Kits. Single-cell suspensions were loaded on a Chromium Single Cell Controller Instrument (10X Genomics) to generate single-cell gel beads in emulsions (GEMs). Briefly, about 16,00020.000 cells were added to each channel, with a targeted cell recovery estimate of 5,0008,000 cells. After generation of GEMs, single-cell RNA-seq libraries were prepared using the Chromium Single Cell 3Library& Cell Bead Kit (10X Genomics) according to the manufacturers protocol. Libraries were sequenced with an IlluminaNovaseq6000 using high-output 75-cycle kits with apreviously reported read length configuration (21 (link)).