title: GSM8369879: Newly diagnosed multiple myeloma _2; Homo sapiens; RNA-Seq
library_construction_protocol: Cryopreserved BM mononuclear cells were rapidly thawed, washed twice with PBS, stained with cell viability dyes and counted using a fluorescence microscope. Samples with viability higher than 95% were processed for 10x genomics scRNA-seq. Approximately 10,000 cells were targeted, and single-cell emulsions were generated with the Chromium Controller (10x Genomics, Pleasanton, CA, USA) using the Chromium Next GEM Chip G (Part Number 1000120, 10x Genomics) with the Chromium NextGEM Single Cell 3' v3.1 kit (Part Number 1000121, 10x Genomics) following the standard protocol. Libraries were assessed for mass concentration using the Qubit 1X dsDNA HS Assay Kit (catalog # Q33231; Invitrogen, Waltham MA, USA) and the Qubit 4 fluorometer (catalog # Q33238; Invitrogen, Waltham MA, USA). Library fragment size was assessed with the High Sensitivity NGS Fragment Analysis Kit (catalog # DNF-474-0500; Agilent, Santa Clara CA, USA) on the Fragment Analyzer System (catalog # M5310AA; Agilent, Santa Clara CA, USA). Libraries were functionally validated with the KAPA Universal Library Quantification Kit (catalog # 07960140001; Roche, Basel CH). Initial low-pass “surveillance” sequencing was performed on an Illumina NovaSeq SP 100-cycle flow cell and data were assessed using the Cell Ranger Count output.