GSM4569784: PBMC from patient 2 sepsis only; Homo sapiens; RNA-Seq
Sorted cells were loaded into the Single Cell Chip and processed through the 10X controller for droplet generation, resulting in droplets containing a single cell and bead per droplet Chromium Single Cell 3' RNA-seq libraries were prepared for sequencing using standard 10X Genomics protocols scRNA-seq was performed using 10X Genomics Single Cell 3' solution, version 2, according to the manufacturer's instructions with 7,000 cells loaded for each sample (n=4 sepsis only and n=3 sepsis+ARDS). Libraries were sequenced on the HiSeq Platform (Illumina) from Novogene and raw BCL/fastq files were analyzed using Cell Ranger version 3.0.2 (10X Genomics). Low-quality cells were removed if they expressed <3000 genes/cell and had more than 20% mitochondrial and 10% hemoglobin gene expression. Post-quality control, 24,806 cells with mean reads 50,419/cell remained for downstream bioinformatic analyses. Consistent analysis was performed and 3212 cells with 182,364 reads per cell were retained after quality control.