GSM6336773: Patient 1-2, scRNA-seq; Homo sapiens; RNA-Seq
Isolated cells were washed with PBS containing 0.04% bovine serum albumin and filtered through a 40-μm cell strainer. Cell concentration and viability were determined by a Countess II Automated Cell Counter with a final cell concentration of 700-1,200 cells/ul. Single-cell RNA-seq libraries were generated using the Chromium Single Cell 3′ Reagent Kit (10X Genomics). Single-cell suspension was mixed with RT-PCR master mix and loaded together with Single Cell 3′ Gel Beads and Partitioning Oil into a Single Cell 3′ Chip (10X Genomics). The cDNA was amplified and further used to construct 3' gene expression library according to the manufacturer's instructions. The size profiles of pre-amplified cDNA and sequencing libraries were examined by the Agilent High Sensitivity 2100 Systems (Agilent). The scRNA-seq library was sequenced on the Illumina NextSeq 500/550 platform.