title: GSM5077759: 20200706_Allograft_1; Mus musculus; RNA-Seq
library_construction_protocol: Single-cell suspensions from naïve and 24 hours after transplantation mouse lung were prepared as described above for flow cytometry with slight modifications. Allograft was removed and digested with 3 mL dispase (Corning) with DNase I (Sigma), and gently teased using forceps into small (1-2 mm) fragments followed by incubation at room temperature with gentle agitation for 30 minutes. The resulting suspension (in DMEM + 5% FBS) was passed through 70 μm cell strainer, erythrocytes were lysed  andlysed and filtered through 40 μm cell strainers. Cells were counted using AO/PI and Cellometer K2 (Nexcelom), and cell viability exceeded 90%. Single-cell 3' RNA-Seq libraries were prepared using Chromium Single Cell V2 Reagent Kit and Controller (10x Genomics). Libraries were assessed for quality (TapeStation 4200, Agilent) and then sequenced on NextSeq 500 or HiSeq 4000 instruments.
attributes.tag: GEO Accession
attributes.value: GSM5077759