title: GSM4576603: Control None 7 r3; Mus musculus; RNA-Seq
library_construction_protocol: Lungs were dissociated using the gentleMACS protocol; flushed through a 70 μm cell strainer and washed twice with HBSS/0.5% FCS/0.4% EDTA buffer, Lung immune cells were then isolated according to the Lymphocyte Separation Medium protocol  and cell pellets were washed to prepare cells for single cell RNA-seq. Handling of the Chromium Controller and library preparation using the Single Cell 3' Reagent Kits v2 (10x Genomics, Pleasanton, CA) were performed; The Single Cell 3' Chip was loaded aiming to capture 5,000 cells. cDNA from captured immune cells was amplified during cDNA amplification (13 PCR cycles) and libraries were prepared from 110 ng of cDNA. 13 amplification cycles were performed during the final index PCR reaction. An additional and final cleanup step with 1 volume of SPRISelect Beads (Beckman Coulter, Brea, CA) was performed to remove any left-over primers and primer dimers. Libraries were on average 450 bp in length and sequencing was performed on a HiSeq 4000 using a HiSeq 3000/4000 PE Cluster Kit and two 50-cycle SBS kits (Illumina, San Diego, CA). Paired-end sequencing included a 26 bp read 1 (16 bp cell barcode and 10 bp UMI), 8 bp index read and 98 bp read 2 (transcript sequence read).