organism: Homo sapiens
description: Protocols: Peripheral blood (PB) and bone marrow (BM) samples were harvested in EDTA containing tubes and stored at room temperature. Samples were processed within 2h from collection. We lysed whole PB or BM with RBC lysis buffer (Biolegned) according to the manufacturer instruction. Total CD15+ neutrophils were isolated with magnetic bead sorting (CD15 Microbeads, Miltenyi) or by florescence activated cell sorting after gating on total neutrophils as CD3-CD19-CD56-CD34-CD14-CD33+CD15+CD193- cells. Cells were sorted at MoFlo XDP (Beckman Coulter) or FACSAria Fusion (Beckton Dickinson) cell sorters. na We generated scRNA-Seq libraries using the microfluidics-based approach of Chromium Single-Cell Controller (10X Genomics) using the Chromium Single Cell 3' Reagent Kit v3.0 according to the manufacturers' instructions. In each experiment we loaded samples in order to obtain a target cell recovery of 10,000 cells. cDNA amplification was performed using 13 PCR cycles. The concentration of the scRNA-seq libraries was determined using Qubit dsDNA HS Assay Kit at Qubit 3.0 and size distribution was assessed using an Agilent 4200 TapeStation system.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-09-09
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-09-09
attributes.tag: External Id
attributes.value: SAMEA11305870
attributes.tag: INSDC center alias
attributes.value: San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), IRCCS San Raffaele Scientific Institute
attributes.tag: INSDC center name
attributes.value: San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), IRCCS San Raffaele Scientific Institute
attributes.tag: INSDC first public
attributes.value: 2022-09-09T12:17:12Z
attributes.tag: INSDC last update
attributes.value: 2022-09-09T12:17:12Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11188:Sample 6
attributes.tag: age
attributes.value: 37
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell isolation strategy
attributes.value: FACS sorting
attributes.tag: cell_type
attributes.value: neutrophil
attributes.tag: clinical history
attributes.value: G-CSF
attributes.tag: common name
attributes.value: human
attributes.tag: individual
attributes.value: UPN25
attributes.tag: organism part
attributes.value: blood
attributes.tag: sample name
attributes.value: E-MTAB-11188:Sample 6
attributes.tag: sex
attributes.value: male