title: GSM8210057: ATAC_MatN_WT_aIL1B_3; Mus musculus; ATAC-seq
library_construction_protocol: LT-HSCs, MPPs, CMPs and GMPs were enriched by neutrophil depletion. Bone marrow single cell suspensions were incubated with 10μg/ml anti-Ly6G (1A8, BioXcell) before neutrophil depletion using sheep anti-rat magnetic beads (Dynabeads, Life Technologies) and subsequently stained with cell surface markers. For sorting pre-neutrophils and neutrophils from bone marrow, blood and lung, cells were first incubated with 500μg/ml purified mouse IgG+10% FCS (BM) or rat anti-mouse CD16/CD32 antibody (blood and lung), stained with cell surface markers and sorted using gating strategies for HSPC populations or CD45+ CD11b+ Ly6G+ 7-AAD- markers for sequencing of blood and lung neutrophils. Nuclei of the sorted cells were isolated, permeabilized and tagmented by using commercially available Tn5 transposase (Illumina, Nextera DNA Library Preparation Kit). DNA fragments underwent a first 9-cycle PCR amplifications using KAPA HiFi HotStart ReadyMix (7958935001, Roche), and we selected the fragments between 150 and 700 bp in size using with SPRI beads (A63881, Backman Coulter). The size-selected fragments were amplified for a second 9-cycle PCR reactions, and the resulted libraries were purified using SPRI beads.