title: GSM6674176: P7_lib2_cre_pos_epcam; Mus musculus; RNA-Seq
library_construction_protocol: Lungs from all experiments were perfused with ice cold HBSS (Hanks' Balanced Salt Solution), were removed and the whole lung was cut in small pieces and was placed in the enzyme solution (5ml Elastase 4U/ml in HBSS + 200ul DNaseI 2.5mg/ml). Suspension is incubated for 1-1,5 hour in 37 C with gentle rotation. The digestion reaction was terminated by adding 20ml HBSS++ (with 10mM HEPES, 2mM EGTA, 1000 U/ml Penicilin, 0.1 mg/ml Streptomycin, 1% BSA). Cells were filtered through a 70μm filter coated with HBSS++, spun down at 400g for 10 min at 4C and resuspended in 10 ml HBSS++. Library was performed according to the manufacter's instructions (single cell 3' v1 protocol, 10x Genomics). Single cell RNAseq library preparatio was done using standard protocol (10x Genomics). Libraries were sequenced together by time point (3 libraries/NovaSeq 6000 SP flowcell).