title: GSM8145495: Mock, replicate 2, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: To ensure representation of both epithelial and hematopoietic lineage cells for each mouse sample, both right and left lungs  were inflated with a lower volume (0.25mL) dispase with agarose plug followed by digestion and processing into single cell  suspension as described above. Cells were next washed 5 times with 5-10mL of PBS+10%FBS and centrifuged at 500xg for  5', followed by a final wash and centrifugation at 200xg for 10' to remove debris. Dead cells were removed by magnetic  separation using Annexin V kit (STEMCELL Technologies). Using the 10X Genomics 3' CellPlex Kit, cells from each mouse sample were tagged using lipid-conjugated Barcode  oligonucleotides according to manufacturer's instructions. Cells were passed through a 40-μM filter and cell viability was  determined using Cellometer 2000 (all showed >90% viability) prior to loading on 10X Chromium instrument. Following  established techniques using Chromium Single Cell 3' Library V2 kit (10X), libraries were constructed and RNA-seq was  performed on each sample targeting 20,000 reads per cell.