title: GSM8701468: HSPC_TET2_CTRL; Homo sapiens; RNA-Seq
library_construction_protocol: HSPCs samples were prepared using 10X 3' V3.1 and sequenced using the HiSeq4000, for immune progeny samples we used the 10X multiome kit and were sequenced in NovaSeq6000. The concentration and viability of the single cell suspension was measured using trypan blue using Eve Automatic Cell Counter. Approximately 3,000-10,000 cells were loaded on Chromium Chip and partitioned in nanolitre scale droplets using the Chromium Controller and Chromium Next GEM Single Cell Reagents (Chromium Single Cell 3' Reagent Kits User Guide (v3.1)). Within each droplet the cells were lysed, and the RNA was reverse transcribed. All the resulting cDNA within a droplet shared the same cell barcode. Illumina compatible libraries were generated from the cDNA using Chromium Next GEM Single Cell library reagents in accordance with the manufacturer's instructions (10x Genomics, CG000315 Chromium Single Cell 3' Reagent Kits User Guide (v3.1 - Dual Index)). For QC of libraries, we used the Agilent TapeStation and sequenced using the Illumina HiSeq4000. Sequencing read configuration: 28-10-10-90. For the immune progeny, we sorted human hematopoietic cell subsets from humanized mice reconstituted with either TET2WT or TET2Mut HSCs as described in Figure S5A. The concentration and viability of the single nuclei suspension was measured using acridine orange (AO) and propidium iodide (PI) and the Luna-FX7 Automatic Cell Counter. Approximately 15,000-20,000 nuclei were transposed, then loaded on Chromium Chip and partitioned in nanolitre scale droplets using the Chromium Controller and Chromium Next GEM Single Cell Reagents (Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Kits User Guide, User Guide, CG000338). Sequencing read configuration: 28-10-10-90 (GEX). The 10x Barcodes in each library type are used to associate individual reads back to the individual partitions, and thereby, to each individual nucleus. scRNA-Seq 10X  3' Gene Expression and 10X Multiome Gene Expression
identifiers.namespace: GEO
identifiers.id: GSM8701468_r1