Protocols: CCE murine ESCs were cultured on 0.1% gelatin-coated tissue culture plates under feeder free culture conditions at 37°C with 5% CO2 in a humidified incubator. The media composition consists of Dulbecco's modified Eagle's medium (DMEM) high glucose, 15% fetal bovine serum (FBS; Gibco), 1% MEM non-essential amino acids (Sigma-Aldrich), 0.1 mM 2-β mercaptoethanol, 1% L-glutamine (Hyclone) and 1% penicillin/streptomycin (Gibco) and Leukemia inhibitory factor (LIF; R&D systems). Early passage mouse ESCs were transducedwith the lentiviral particles in mouse ESC media supplemented with polybrene (8μg/ml) for 24 hours. After 36 hours, infected cells were treated with puromycin (2μg/ml)for additional 6 days. Extraction of total RNA was performed using RNeasy Plus Mini Kit (Qiagen). One μg of total RNA was used for complementary DNA synthesis using RevertAid First Strand cDNA Synthesis kit (Invitrogen) as per the manufacturer's instructions. RNAseq library preparation was carried out at Novogene facilities (https://en.novogene.com/) following Illumina guidelines.
ENA first public
2021-03-31
ENA last update
2021-02-16
External Id
SAMEA8082543
INSDC center alias
Departamento de Bioquimica y Biologia Molecular y Genetica, Universidad de Extremadura
INSDC center name
Departamento de Bioquimica y Biologia Molecular y Genetica, Universidad de Extremadura
INSDC first public
2021-03-31T04:07:29Z
INSDC last update
2021-02-16T12:11:03Z
INSDC status
public
Submitter Id
E-MTAB-10108:SC1
broker name
ArrayExpress
cell line
CCE
cell type
mouse embryonic stem cell
common name
house mouse
developmental stage
embryo
genotype
wild type genotype
organism part
blastocyst
sample name
E-MTAB-10108:SC1
strain
129S/SvEv-Gpi1<c>