GSM6304118: microglia and myeloid cells multiplexed scRNA-seq rep2, GEX; Homo sapiens; RNA-Seq
GSM6304118
Multiplexed single-cell sequencing with cell hashing was performed following previously published protocol (30567574). In brief, each type of cells were first blocked with Human TruStain FcX (BioLegend Catalog# 422302) in cell staining buffer (BioLegend Catalog# 420201) for 10 minutes on ice, followed by staining with an individual TotalSeq-A anti-human Hashtag for 45 minutes on ice. Cells were washed three times, resuspended in PBS with 0.04% bovine serum albumin, and counted with Countess II Automated Cell Counter (Thermo Fisher Scientific). 1.25 x 10^5 cells were pooled from each group to create a pool of 1x10^6 cells with an average viability of 85%. Pooled cells were then filtered through a 40 μm filter, counted, and diluted to a final concentration of 730 cells/μL. Single cell suspension containing 6.4x105 cells were loaded into 10X Chromium Controller following manufacturer's user guide (Document CG000185 Rev B) using 2 lanes of Chromium Single Cell 3' Reagent Kits v3. Single cell gene expression and hashtag barcode libraries were constructed following the manufacturer's user guide (Document CG000185 Rev B). Final libraries were sequenced on Illumina NovaSeq 6000 sequencer using NovaSeq 6000 S4 Reagent Kit v1 to get a total of 2x109 reads.