title: GSM8875908: Tonsil B-cells, Right Gene Expression; Homo sapiens; RNA-Seq
library_construction_protocol: Cryopreserved human tonsil cells from 4 Strep A-colonised paediatric donors were thawed, stained with GAC and SpyCEP antigen tetramers separately, and magnetically enriched for tetramer-binding cells. These cells were stained with surface marker antibodies and either TotalSeq™-C0252 anti-human Hashtag 2 Antibody (for SpyCEP tetramer-binding cells) or TotalSeq™-C0254 anti-human Hashtag 4 Antibody (for GAC tetramer-binding cells). Antigen tetramer-binding isotype switched (IgD–) B cells were isolated by Fluorescence-activated cell sorting (FACS) and sequenced by single cell sequencing using the Chromium Next GEM Chip K Single Cell Kit (10X Genomics, PN-1000286), the Chromium Next GEM single Cell 5' v2 dual index kit (10X Genomics, PN-1000263). Sequencing libraries were generated following the Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) with Feature Barcode technology for Cell Surface Protein & Immune Receptor Mapping protocol (10X Genomics, CG000330 Rev C). Single cell RNA, VDJ, and feature barcode sequencing libraries were generated using the Chromium Next GEM Chip K Single Cell Kit (10X Genomics, PN-1000286), the Chromium Next GEM single Cell 5' v2 dual index kit (10X Genomics, PN-1000263), the 10X Chromium Single Cell Human BCR Amplification Kit (10X Genomics, PN-1000253), and the 5' Feature Barcode Kit (10X Genomics, PN-1000256). Libraries were indexed using the Dual Index Kit TT Set A (10X Genomics, PN-1000215) and the Dual Index Kit TN Set A (10X Genomics, PN-1000250), pooled for sequencing at a molar ratio of 3:1:1 (GEX:VDJ:feature barcode), and sequenced with the Illumina Novaseq 6000 on an S1 flowcell. 10X single cell RNAseq sequneced for GEX, VDJ and Feature barcoding. Cells were spread across two lanes (L= left, R = right) and were further spread across two lanes, L001 and L002.
identifiers.namespace: GEO
identifiers.id: GSM8875908_r1