GSM5734076: LL17LL; Rattus norvegicus; RNA-Seq
Lungs were extracted from the animals and transferred into 60mm petri dish. A syringe fitted with 25G needle was injected into lung pieces with enzyme solution (DMEM (low glucose) + 1 mg/mL Collagenase/Dispase + 3 U/mL Elastase + 20 U/mL DNAse).  The tissue pieces were minces into small pieces and transfered to a 50 mL conical tube. Incubate the tissue pieces at 37C for 20 minutes at a agitation of ~75rpm. The tissue was dissociated through a wire mesh strainer and rinsed with medium, followed by centrifugation and filtration. The number of cells was counted and the cell solution was diluted to a final concentration of 1M cells/mL in 0.01% BSA in PBS. scRNAseq libraries were generated with the Chromium Single Cell 3' assay (10x Genomics). Samples were diluted for an expected cell recovery population of 10,000 cells per lane. Bulk RNAseq libraries were prepared using the Pair End Sequencing Prep Kit. For scRNAseq, libraries sequenced on the HiSeq4000 platform to a depth of 180 to 500 million reads per library. Downstream processing was conducted with Cellranger 3.0.2 with the default parameters. For bulk RNAseq, libraries were sequenced through NovaSeq with sequencing read length of 100 base pairs (bp).