title: GSM4151705: adult murine lung GLI1+ stromal cells treated with bleomycin; Mus musculus; RNA-Seq
library_construction_protocol: Whole lung was dissected from the adult mouse and tracheally perfused with a digestion cocktail of Collagenase Type I (225 U/ml, Thermo Fisher), Dispase (15 U/ml, Thermo Fisher) and Dnase (50 U/ml, Sigma) and removed from the chest. The lung was further diced with razor blades and the mixture was incubated for 45 mins at 37 °C and vortexed continuously. The mixture was then washed with sorting buffer (2% FBS, 1% Pen-Strep in DMEM). The mixture was passed through a 70 µm cell strainer and resuspended in RBC lysis buffer, before passing through a 40 µm cell strainer. Cell suspensions were resuspended in sorting buffer. DAPI (0.2 µg/ml) was used to exclude dead cells. Doublets and dead cells were excluded based on forward and side scatter and DAPI fluorescence respectively. Cells were sorted into sorting buffer. FACS analysis was performed by FACSDiva. This preparation was performed separately for each sample. cDNA libraries were prepared for sequencing using standard protocol for the 10x Single Cell instrument. The libraries for the two samples were prepared and sequenced separately. Droplet-based single cell RNA-Seq
attributes.tag: GEO Accession
attributes.value: GSM4151705