title: Illumina HiSeq 2500 sequencing; Transcriptomic characterization of classical monocytes highlights the involvement of immuno-inflammation in bone erosion in Rheumatoid Arthritis
design: Transcriptomic characterization of classical monocytes highlights the involvement of immuno-inflammation in bone erosion in Rheumatoid Arthritis
library_construction_protocol: Blood (70 ml) from RA patients and control group were collected, and peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll density and dextran sedimentation. In both studied groups, monocytes were isolated from PBMCs by depletion of non-monocytes cells using an indirect magnetic labeling system. Classical monocytes were indirectly isolated using the CD16 MicroBeads human kit (Miltenyi Biotec®, Germany). Classical monocytes RNA was isolated using the RNeasy Plus Mini (Qiagen®), and RNA integrity number (RIN ≥ 7) was evaluated by the ScreenTape method using Bioanalyser 2100 (Agilent®). RNA library was constructed using the Quant-seq® 30 mRNA-Seq Library Prep Kit (Lexogen®).