alias: GSM4544005
title: GSM4544005: NL_1; Homo sapiens; RNA-Seq
library_construction_protocol: Human lung fragments were digested with a cocktail of Collagenase Type I (225 U/ml, Thermo Fisher), Dispase (15 U/ml, Thermo Fisher) and Dnase (50 U/ml, Sigma) . The cell suspension then washed with FACS buffer (2% FBS in DMEM-F12). The mixture was passed through a 70 µm cell strainer and resuspended in RBC lysis buffer, before passing through a 40 µm cell strainer. Cells suspensions were incubated with the appropriate antibodies in FACS buffer for 30 min at 4 °C and washed with FACS buffer. The following antibodies were used at 1:200 for staining: anti-CD45, anti-CD11b, anti-CD31, and anti-Epcam. DRAQ7 was used to exclude dead cells. Doublets and dead cells were excluded based on forward scatter, side scatter and DRAQ7 fluorescence. Cells were sorted into FACS buffer. FACS analysis was performed by FACSDiva. cDNA libraries were prepared for sequencing using standard protocol for the 10x Single Cell instrument, using Single cell 3' v2 chemistry kit.
identifiers.namespace: GEO
identifiers.id: GSM4544005