GSM9009681: MCD, scRNA-seq, rep 2; Mus musculus; RNA-Seq
The scRNA-Seq libraries were performed using the 10X Genomics Chromium Controller Instrument and Chromium Next GEM Single Cell 30 Reagent Kits v3.1. In brief, mouse spleens were detached from euthanized mice and mashed through the 40 μm nylon cell strainer (BD Falcon) for splenocytes suspension. Cells were quality controlled to ensure > 90% viability, washed and resuspended to approximately 1000 cells/μL and loaded into appropriate channels to generate single-cell Gel Bead-In-Emulsions (GEMs), and the cells were lysed to release mRNA and reverse transcribed to cDNA. Thereafter, the GEMs were broken, and the barcoded cDNA was purified, amplified and fragmented, A-tailed to ligate with adapters and index PCR amplified. The constructed libraries were quantified by Qubit High Sensitivity DNA assay and the size distribution was checked using a High Sensitivity DNA chip in the Bioanalyzer 2200 (Agilent). All libraries were sequenced by Illunima Novaseq 6000 platform at Biotechnology Corporation (Shanghai, China).