Patient PDX osteosarcoma tibia tumor single nuclei RNAseq - NCH-OS-4
Tissues were processed using the human tumor dissociation kit (Miltenyi Biotec, 130-095-929) with a gentleMACS Octo Dissociator with Heaters (Miltenyi Biotec, 130-096-427) for 3′ single-cell analysis. For single-nucleus analysis, cryopreserved samples were isolated and permeabilized for single-cell gene expression sequencing following the Nuclei Isolation from Complex Tissues for Single Cell Multiome ATAC + Gene Expression Sequencing protocol (10x Genomics, CG000375 Rev C). Following dissociation into single cells or nuclei, 3′ RNA-seq libraries were prepared using Chromium Single Cell 3′ RNA-seq system (10x Genomics) with the Reagent Kit v3.1 (10x Genomics, PN-1000121) according to the manufacturer's instructions. cDNA library quality and quantity were determined using an Agilent 2100 bioanalyzer using the High Sensitivity DNA Reagents kit (Agilent, 5607-4626) and then sequenced on an Illumina NovaSeq 6000.