characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2019-11-05T04:04:27Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2019-10-15T16:22:50Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA6086357
characteristics.characteristic: External Id
characteristics.text: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas
characteristics.characteristic: INSDC center name
characteristics.text: 2019-11-05T04:04:27Z
characteristics.characteristic: INSDC first public
characteristics.text: 2019-10-15T16:22:50Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS3886765
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-8439:Ercc1_KO_Rep1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: bone marrow macrophage
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002476
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Er1F/+ and Er1F/- BMDMs (bone marrow derived macrophages)were differentiated from bone marrow precursors. Briefly, bone marrow cells were isolated from mouse femurs and tibias and cultured for 7 days in DMEM containing 10% FBS, 30% L929 conditioned media, 50μg/ml streptomycin, 50 U/ml penicillin (Sigma) and 2mM L glutamine (Gibco). Total RNA was isolated from Er1F/+ and Er1F/- BMDMs (bone marrow derived macrophages) using a Total RNA isolation kit (Qiagen) as described by the manufacturer. Library preparation started with 1µg total RNA. After poly-A selection (using poly-T oligo-attached magnetic beads), mRNA was purified and fragmented using divalent cations under elevated temperature. The RNA fragments underwent reverse transcription using random primers. This is followed by second strand cDNA synthesis with DNA polymerase I and RNase H. After end repair and A-tailing, indexing adapters were ligated. The products were then purified and amplified to create the final cDNA libraries. After library validation and quantification (Agilent 2100 Bioanalyzer), equimolar amounts of all 12 libraries were pooled. The pool was quantified by using the Peqlab KAPA Library Quantification Kit and the Applied Biosystems 7900HT Sequence Detection System.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: Er1F/-
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: bone marrow
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: mixed
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001271
characteristics.characteristic: sex
characteristics.text: C57BL/6
characteristics.characteristic: strain
characteristics.text: Ercc1_KO_Rep1
characteristics.characteristic: title