title: GSM7317507: 15M CD-HFD (Low AFP); Mus musculus; RNA-Seq
library_construction_protocol: The liver of diet-fed mice were perfused with collagenase and dissociated to separate out parenchymal (hepatocytes, cholangiocytes) and non-parenchymal (endothelial cells, immune cells etc..) fractions. Parenchymal fractions were purified using a 30% percoll gradient. Non-parenchymal fraction were enriched for live cells by flourescent activated cell sorting. Purified parenchymal and non-parenchymal cells were combined at a 1:1 ratio before submission for sequencing. Single cells were encapsulated into droplets and processed following standard manufacturer's specification, using 10X Genomics GemCode technology. Cells were loaded on a 10x Genomics Chromium controller instrument to generate single-cell Gel Beads in emulsion (GEMS) at the Yale Center for Genome Analysis. Lysis and barcoded reverse transcription of polyadenylated mRNA from single cells were performed inside each GEM followed by complementary DNA (cDNA) generation using the single-Cell 3' Regent Kits version 2 (10X Genomics). Libraries were sequenced on an Illumina HiSeq 4000 as 2x 100 paired-end reads.