GSM8927782: ratlungN; Rattus norvegicus; RNA-Seq
Freshly obtained rat whole lungs were finely chopped before treatment with 0.1 mg/mL collagenase-D in DMEM and 100ug/mL DNase-I for 1 h. The treated tissue was mashed through a 100 µm cell strainer and washed with 20 mL DMEM containing 10% FBS and 1% antibiotics. The cell suspension was centrifuged for 5 min at 300g to pellet the cells. Red cells were removed by the lysis buffer at room temperature for 60 seconds. The suspension was then serially washed with PBS containing 0.01% BSA before centrifuging at 300 g for 5 min and went through a 70 µm cell strainer once and 40 µm cell strainers twice. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics)