title: GSM3142409: OCP_BOTH_5_run2: OCP_RNASeq - Osteoclast_precursors Combined_treatment replicate_5 run_2; Mus musculus; RNA-Seq
library_construction_protocol: RNA isolated from mouse bone marrow was subject to quality control using the Bioanalyzer (Agilent). BMCs were prepared for flow cytometry by suspension in sterile PBS containing 2% FBS. Cells were labeled with appropriate antibodies for 30 minutes at 4°C. Gating was used to exclude debris, cell clumps, and dead cells (using 7-aminoactinomycin D; 7-AAD Viability Staining Solution (BioLegend, 420404).Prior to sorting of the M/OCP populations, the Murine Direct Lineage Cell Depletion Kit (Miltenyi Biotec Inc., 130-110-470) was used to enrich for Lin- populations, which was confirmed by flow cytometry for the markers in the lineage cocktail. Myeloid/osteoclast progenitor cell (M/OCP) populations were defined as Lineage- CD115+.  Antibody panel includes: Pacific BlueTM anti-mouse Lineage Cocktail (BioLegend, 133310: CD3-, Ly-6G/Ly6-C-, Cd11b-, CD45R-, TER119-) and Alexa Fluor® 488 anti-mouse CD115 (CSF-1R; BioLegend, 135511). RNA was converted into a DNA library using the Ultra RNA Library Prep Kit for Illumina (New England BioLabs). After further quality control, libraries were sequenced on the HiSeq 2000 (Illumina).
attributes.tag: GEO Accession
attributes.value: GSM3142409