title: Sample 3
organism: Homo sapiens
description: Protocols: Blood samples were collected from 3 healthy volunteers and peripheral blood mononuclear cells (PBMCs) isolated by density gradient centrifugation on Ficoll. PBMCs were differentiated into macrophages using macrophage colony-stimulating factor (M-CSF). Cells were stimulated for 3h as detailed and total RNA extracted. PBMCs were differentiated to macrophages in RPMI + 10% FCS with 50ng/ml M-CSF for 8 days before stimulation. PBMC-derived macrophages were stimulated with either media alone, 200ng/ml lipid A or 50ng/ml TNF alpha for 3h before RNA isolation. ~2x10^6 cells were stimulated per condition. RNA was extracted using the mirVana RNA isolation kit according to the manufacturer's instructions. Input RNA quality was assessed on a TapeStation2200 using the RNA Screen Tape to confirm high RNA quality (RIN score > 8) according to manufacturers instructions. Input RNA quantity was assessed using a Qubit 2.0 Fluorometer and the Qubit RNA BR Assay Kit according to manufacturers instructions. Library was constructed using the Illumina TruSeq stranded total RNA kit according to manufacturers instructions with an input of 500 ng total RNA / sample. This kit includes ribosomal RNA removal by Ribozero treatment. The quality of final libraries was checked on a TapeStation2200 using the D1000 Screen Tape.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-12-31T17:02:25Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-23T09:50:05Z
attributes.tag: External Id
attributes.value: SAMEA104717290
attributes.tag: INSDC center name
attributes.value: Warwick Medical School, University of Warwick, UK
attributes.tag: INSDC first public
attributes.value: 2018-12-31T17:02:25Z
attributes.tag: INSDC last update
attributes.value: 2018-03-23T09:50:05Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6607:Sample 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: macrophage
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: individual
attributes.value: 1
attributes.tag: organism part
attributes.value: blood
attributes.tag: progenitor cell type
attributes.value: peripheral blood mononuclear cell
attributes.tag: sample name
attributes.value: E-MTAB-6607:Sample 3
attributes.tag: scientific_name
attributes.value: Homo sapiens