GSM7244779: M1 HSPCs, barcode-derived cDNA; Homo sapiens; OTHER
We performed DAbseq on unsorted mononuclear cells from 14 patients using a microfluidic approach with molecular barcode technology using the Tapestri platform (MissionBio) as previously described . Briefly, cryopreserved cells were blocked and stained with a pool of 22 oligo-conjugated antibodies. Individual samples were also incubated with unique anti-CD45 oligo-conjugated antibodies to provide sample-level identifiers, and groups of 2-3 individual patients were pooled together for multiplexed runs. Pooled samples were run through Mission Bio Tapestri platform as previously described. DNA from barcoded cells was amplified via PCR using a targeted panel that included 127 amplicons across 20 genes associated with acute leukemia. DNA PCR products were isolated and purified with standard library preparation protocol. Protein PCR products were isolated from the supernatant from AmpureXP bead purification via incubation with a 5' Biotin Oligo (ITD), and purified using Streptavidin C1 beads.