GSM9114532: NPCs, liver, C.sinensis 16 wpi, GEX, Rep2; Mus musculus; RNA-Seq
Liver dissociation was performed by Liver Dissociation Kit (Miltenyi Biotec, 130-105-807) according to the manufacturer's instructions. Then the liver single cell suspension was subjected to density centrifugation using 30% Histodenz-HBSS (Sigma-Aldrich, D2158) gradient centrifugation as described. In brief, suspended the cells with 30% Histodenz, slowly added some HBSS on it and centrifuged at 1400 g for 20 min at 4 °C without braking. Cells at the interface were enriched liver non-parenchymal cells (NPCs). Libraries were prepared with the 10x Genomics Chromium Single Cell 5′ Gene Expression chemistry following the manufacturer's user guide. Cells were encapsulated in GEMs, where poly-A RNA was reverse-transcribed to barcoded cDNA. Barcoded cDNA was purified, PCR-amplified, size-selected with SPRIselect, and finalized as paired-end sequencing libraries. T cell receptor libraries were constructed with the 10x Genomics Chromium Single Cell V(D)J chemistry according to the official protocol. Barcoded full-length cDNA generated in GEMs underwent two rounds of targeted PCR to enrich TCR variable regions, followed by purification and indexing to yield paired-end sequencing libraries.
GEO
GSM9114532_r1