title: GSM8804808: Sed_liver_2; Mus musculus; RNA-Seq
library_construction_protocol: Sed and PEx mice bearing MC38 tumors were sacrificed 3 weeks after tumor injection. The liver was harvested and processed into single-cell suspensions using liver dissociation kits and then passed through a 70 μm cell strainer. Differential centrifugation separated non-parenchymal cells (NPCs) from hepatocytes (50 g for 5 minutes, repeated twice). The supernatant was centrifuged again (300 g for 10 minutes) to obtain CD45+ immune cells were sorted using CD45+ beads from these single-cell suspensions, achieving a purity greater than 90%. Following quality control checks for cell viability and concentration, samples were transferred to a pre-chilled 96-well plate, heat-sealed, and reverse transcription was carried out using a Veriti 96-well thermal cycler. Following reverse transcription, cDNA was extracted using Recovery Agent (10x) and purified with a Silane DynaBead clean-up as described in the user guide. The purified cDNA was then amplified for 12 cycles, diluted at a 4:1 ratio (elution buffer, Qiagen, to cDNA), and analyzed on a Bioanalyzer (Agilent Technologies) to measure cDNA concentration. cDNA libraries were prepared using the 10X Chromium Single Cell 3' Library and Gel Bead Kit v3.1.
identifiers.namespace: GEO
identifiers.id: GSM8804808_r1