GSM8366583: CD45 negative live cells  from bone metastatic tumor; Mus musculus; RNA-Seq
Single cell suspension samples were examined on Attune flow cytometer for cell density and viability. For each sample, 15,000 sorted cells were resuspended in 15 ul of PBS containing 0.5% BSA for library preparation. Around 15,000 cells from each sample were loaded into each channel of the 10X Genomics Chromium X system using the Chromium Single Cell 3' v3.1 Reagent Kits (10X Genomics Inc., CA) to generate and amplify cDNA from individual cells. The amplified cDNA samples were purified with Ampure XP magnetic beads (Beckman Coulter, CA), quantified by Qubit fluorometer (Invitrogen, CA), and examined on Bioanalyzer with High Sensitivity DNA chips (Agilent, CA) for size distribution. Illumina sequencing libraries were generated from the amplified cDNA samples using the Illumina Tagment DNA Enzyme and Buffer kit (Illumina, CA). These libraries were examined by Qubit and Bioanalyzer, then pooled at equal molar amount and sequenced on Illumina NovaSeq 6000 S Prime flowcells as 28+94 nt pair-end reads following the standard Illumina protocol.