FACSorting of neutrophils from total BM samples of MM patients and HA Approximately 20,000 cells of various neutrophils subsets were FACSorted from total BM samples of HA (N = 3) or MM patients (N = 3), and placed in a PCR tube with 100µL of PBS+BSA 0.05%. After centrifugation (500g, 4ºC, 10 min), the pellet was suspended in 25µL of the transposase reaction mixture (15µL of Buffer TD2x (Illumina), 1µL of TDE1 enzyme (Illumina), 0.25µL of 5%digitonin (Promega) and 8.75µL of nuclease-free water). The resulting mix was incubated at 37ºC for 30 min at 450 rpm. Afterwards, the transposase reaction was stopped on ice and 5µL of clean up buffer, 2µL of 5% SDS and 2µL of Proteinase K (New England Biolabs) were added to the previous mix and incubated for 30 min at 40ºC. Transposase-reaction products were cleaned up with AMPure magnetic beads 2X (BC). Finally, the DNA fragments were amplified with 22µL Kappa Hifi ready mix + 4µL of primer 1 i5 and 2 i7 mix per sample. Library was purified with AMPure magnetic beads 2X. Quality control was performed with Qubit and TapeStation
GEO Accession
GSM4520712