characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-11-30T00:14:48Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-11-30T00:14:48Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA110669171
characteristics: External Id
characteristics: Novo Nordisk Foundation Center for Stem Cell Biology (DanStem). University of Copenhagen.
characteristics: INSDC center name
characteristics: 2022-11-30T00:14:48Z
characteristics: INSDC first public
characteristics: 2022-11-30T00:14:48Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS12766750
characteristics: SRA accession
characteristics: E-MTAB-12104:CM WT R1
characteristics: Submitter Id
characteristics: ArrayExpress
characteristics: broker name
characteristics: epiblast stem cells
characteristics: cell_line
characteristics: cardiac mesoderm
characteristics: cell_type
characteristics: house mouse
characteristics: common name
characteristics: Protocols: Cells were seeded on fibronectin-coated 6-well plates in N2B27 medium (DMEM-F12:Neurobasal, 1:1, N2, B27) supplemented with Activin A, FGF and XAV-939 and differentiated to all desired cell types as described in the publication. At 24h (EpiSC, PPS and APS) and 48h (mesoderm) cells were washed briefly and incubated with accutase for 3 min at 37C. After that time accutase was diluted with 3x volume N2B27, samples were centrifuged and the cell pellet was resuspended in lysis buffer (QIAGEN RNAeasy kit). RNA was extracted using QIAGEN RNAeasy kit according to manufacturer's indications. RNA concentration was quantified by NanoDrop and quality was verified with a Fragment analyzer. Library preparation was carried out using 0.5 μg of RNA with NEBNext Ultra II RNA library Prep Kit (New England BioLabs, E7770S) as per manufacturer’s instructions. Libraries were amplified for 4 total PCR cycles and purified with AMPure XP beads. DNA concentration was measured with a Qubit Fluorometer and fragment size was determined with a Fragment analyzer.
characteristics: description
characteristics: embryo
characteristics: developmental stage
characteristics: normal
characteristics: disease
characteristics: wild type genotype
characteristics: genotype
characteristics: Mus musculus
characteristics: organism
characteristics: Mus musculus
characteristics: organism
characteristics: epiblast
characteristics: organism part
characteristics: pluripotent stem cell
characteristics: progenitor cell type
characteristics: Mus musculus
characteristics: scientific_name
characteristics: CM WT R1
characteristics: title