title: GSM8791438: AML Patient 28830; Homo sapiens; RNA-Seq
library_construction_protocol: Frozen mononuclear cells from AML samples were thawed, counted and enriched for live cells by depletion of dead cells (Miltenyi Biotec, catalog number 130-090-101). Isolated live cells were washed three times with 1X Phosphate-Buffered Saline (PBS, calcium and magnesium free) containing 0.04% weight/volume BSA (Thermo Fisher Scientific, catalog number AM2616) and automatedly counted by the Countess 3 Automated Cell Counter (Thermo Fisher Scientific) and manually by the Neubauer hemocytometer. From each sample, we calculated the volume of cells to load to have a desired recovery target of 8,000-10,000 cells and loaded on Chromium Next GEM Chip G (10X Genomics; PN-2000177) with reagents from Chromium Next GEM Single Cell 5' Library & Gel Bead Kit v1.1 (10X Genomics; PN-1000165), following standard manufacturer's protocols for gel beads-in-emulsion (GEMs) generation, barcoding, cDNA amplification and gene expression library construction. Final library quality was assessed using High Sensitivity D1000 ScreenTape (5067- 5584) with High Sensitivity D1000 Reagents (5067- 5585) at the Agilent 2200 TapeStation system. Illumina-ready dual index libraries were sequenced at the recommended depth and aiming at 50,000 reads/cell at the Hartwell Center at St Jude Children's Research Hospital (SJCRH) on the Illumina NovaSeq according to manufacturer's recommendations.