title: GSM6634890: STAT6_post_T_GEX; Homo sapiens; RNA-Seq
library_name: GSM6634890
library_construction_protocol: scaling and clustering was performed on PBMCs and enriched T cells separately.  Dimensionality reduction using PCA was done on the 2000 most variable genes and UMAP was based on the first 20 PCs. Single cell encapsulation and library preparation are done at Genomics Core of the Centre for PanorOmic Sciences in LKS Faculty of Medicine of The University of Hong Kong. Single cell encapsulation and cDNA libraries were prepared by Chromium™ Single Cell 5' Reagent Kit v1 and Chromium™ Single Cell A Chip Kit, or by Chromium Next GEM Single Cell 5' Reagent Kit v2 (Dual Index) and Chromium Next GEM Chip K Single Cell Kit. Cells in suspension were counted and loaded into individual wells of 10X Chromium Single Cell chip (10X Genomics, Pleasanton, United States). Single cells were then encapsulated into Gel Beads-in-emulsion (GEM) by 10X Chromium Single Cell Controller. Single Cell 5' Reagent Kit was used to perform downstream steps: reverse transcription is performed on GEMs, followed by cDNA clean-up and amplification. The double-stranded cDNA will then go through enzymatic fragmentation, adapter ligation, index PCR and SPRIselect size selection was done as per manufacturer's protocol. Library size and concentration are determined by Qubit, quantitative PCR and Bioanalyzer assays. The libraries were denatured and diluted to optimal concentration. Illumina NovaSeq 6000 was used for pair-end 151bp sequencing with NovaSeq 6000 S4 Reagent Kit v1.0 and v1.5 (Illumina #20012866 and #20028312, San Diego, US).