Mus musculus
Protocols: Naive murine embryonic stem cells (mESC) were derived freshly (FVB or mixed 129/B6) and were routinely cultured on gelatin-coated plates in t2i/L media: NDiff (N2B27) (Takara #y40002) supplemented with titrated 2i (0.2 �M PD0325901 and 3 �M CHIR 99021), 1,000 U/ml leukaemia inhibitory factor (LIF), 1% FBS, 1% penicillin streptomycin and maintained in humidified atmosphere at 37C and 5% CO2. Cells were passaged every 2 days by dissociation with TrypLE and medium was changed daily. To induce epiblast-like cell (EpiLC) differentiation, naive mouse ESCs were seeded on fibronectin-coated plates in NDiff (N2B27) (Takara #y40002) supplemented with 20 ng/ml ActivinA (PeproTech #120-14P), 12 ng/ml bFGF (PeproTech #450-33), 1% knockout serum replacement (Thermo Fisher #10828010) and 1% penicillin streptomycin. Cell were harvested ~40 hours after EpiLC induction. RNA-sequencing of polyA RNA from mouse Embryonic Stem Cells. Total RNA was isolated using the Monarch Total RNA Miniprep Kit (NEB #T2010S), including gDNA removal step and on-column DNase I digestion. Total RNA was quantified using Qubit II and quality assessed with high-sensitivity RNA Screen Tape (Agilent 5067-5579). Stranded mRNA-seq libraries were prepared with the NEBNext RNA Ultra II kit (input: 350 ng of total RNA) following the manufacturer's instructions.
ENA-FIRST-PUBLIC
2023-11-15T10:00:25Z
ENA-LAST-UPDATE
2023-11-15T10:00:25Z
External Id
SAMEA114497937
INSDC center name
EMBL
INSDC first public
2023-11-15T10:00:25Z
INSDC last update
2023-11-15T10:00:25Z
INSDC status
public
Submitter Id
E-MTAB-13467:mll2wt_EpiLC_rep2
broker name
ArrayExpress
cell_line
mESC
cell_type
EpiLC
collection date
not collected
common name
house mouse
genetic modification
targeted mutation
genotype
wild type genotype
geographic location (country and/or sea)
not collected
isolate
not applicable
sample name
E-MTAB-13467:mll2wt_EpiLC_rep2
scientific_name
Mus musculus
sex
female
strain
FVB