WT_Rep1
Mus musculus
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.
ENA-FIRST-PUBLIC
2019-11-05T04:04:27Z
ENA-LAST-UPDATE
2019-10-15T16:23:00Z
External Id
SAMEA6086360
INSDC center name
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas
INSDC first public
2019-11-05T04:04:27Z
INSDC last update
2019-10-15T16:23:00Z
INSDC status
public
Submitter Id
E-MTAB-8439:WT_Rep1
broker name
ArrayExpress
cell type
bone marrow macrophage
common name
house mouse
developmental stage
adult
genotype
wild type genotype
organism part
bone marrow
sample name
E-MTAB-8439:WT_Rep1
scientific_name
Mus musculus
sex
mixed
strain
C57BL/6