GSM6447689: LTB5109, CMML3; Homo sapiens; RNA-Seq
60 million cryopreserved BM-MNCs per sample were obtained. Samples were thawed, washed with 20% FBS in PBS and treated with DNAse I for 15min to reduce cell clumping. First, dead cell removal was performed according to manufacturer instructions (Miltenyi Biotech, Catalog number: 130-090-101) followed by CD34 enrichment, also according to manufacturer protocol (Miltenyi Biotec, Catalog number: 130-046-703). Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics). Briefly, Chromium Single Cell 3' Library, Gel Bead & Multiplex Kit and Chip Kit (10X Genomics v3) was used to encapsulate and barcode for cDNA preparation of enriched CD34+ BM-MNCs in 40 samples across 29 unique patients. The targeted cell population sizes were 5,000 cells for each sample (Supplementary Table S7). The libraries were constructed according to the manufacturer's protocol, sequenced on an Illumina NovaSeq, and mapped to the human genome (GRCh38) using CellRanger (10X Genomics).