AML-122-001
Simultaneous profiling of DNA mutation and cell-surface immunophenotype was performed according to the Mission Bios protocol using the custom-designed panel kit and 10 to 15 oligo-conjugated antibodies (CD13, CD33, CD3, CD19, CD22, CD11b, CD14, CD64, CD34, HLA-DR, CD117, CD123, CD38, CD90, and CD45) purchased from Mission Bio, Inc. Briefly, cryopreserved bone marrow mononuclear cells were thawed, quantified and then stained with the pool of 10 to 15 oligo-conjugated antibodies. The stained cells were washed and loaded onto the Tapestri  machine for single-cell encapsulation, lysis, and barcoding, following the protocol similar to the scDNA-seq except for adding an extra primer for antibody tags prior to the barcoding. The barcoded samples were then thermocycled. DNA libraries was extracted from the droplets followed by the purification using Ampure XP beads (Beckman Coulter). The supernatant from Ampure XP beads incubation contained antibody-tagged libraries, and was incubated with biotinated oligo (Integrated DNA Technologies) to capture the antibody tags, followed by the purification using streptavidin beads (Thermo Fisher Scientific). The purified DNA and antibody-tagged libraries were indexed and then sequenced on Illuminas NovaSeq 6000 or NextSeq 500 systems with 150- or 250-bp paired-end multiplexed runs.