title: GSM4996760: Hepatic γδ T lymphocytes; Mus musculus; RNA-Seq
library_construction_protocol: Using single cell 3 'Library and Gel Bead Kit V2 (10x Genomics, 120237) and Chromium Single Cell A Chip Kit (10x Genomics, 120236), the cell suspension (300-600 living cells per microliter determined by Count Star) was loaded onto the Chromium single cell controller (10x Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer's protocol. In short, single cells were suspended in PBS containing 0.04% BSA. These cells were added to each channel, and the target cell will be recovered was estimated to be about 8600 cells for sample 1 and 4500 cells for sample 2. Captured cells were lysed and the released RNA were barcoded through reverse transcription in individual GEMs. Reverse transcription was performed on a S1000TM Touch Thermal Cycler (Bio Rad) at 53°C for 45 min, followed by 85°C for 5 min, and hold at 4°C. The cDNA was generated and then amplified, and quality assessed using an Agilent 4200 (performed by CapitalBio Technology, Beijing). According to the manufacture's introduction, Single-cell RNA-seq libraries were constructed using Single Cell 3' Library Gel Bead Kit V2. The libraries were finally sequenced using an Illumina Novaseq 6000 sequencer with a sequencing depth of at least 45,000 reads per cell with pair-end 150 bp (PE150) reading strategy (performed by CapitalBio Technology, Beijing).