ZFC4mut_1
Mus musculus
Protocols: Cells were directly lysed on the plate with RLT lysis buffer (Qiagen RNeasy Plus Mini Kit, cat. 74134). The material was transferred to microcentrifuge tubes and stored at -80°C before RNA extraction. ESC lines were grown in gelatinized dishes in Glasgow MEM (Gibco, UK) supplemented with 15% fetal bovine serum (Hyclone; ThermoFisher Scientific Scientific, UK), 1% sodium pyruvate, 1% non-essential amino acids, 0.1% β-mercaptoethanol and leukemia inhibitory factor (LIF). For RNA-seq, ESC lines were grown for 7-10 days to obtain a healthy, fast growing culture. Then 6x105 cells were seeded into wells of 6-well plates and were grown for 24h before sample collection. For each cell line, four independent cell seedings were performed to obtain replicates. Total RNA was extracted using the Qiagen RNeasy Plus Mini Kit (cat. 74134), following manufacturer's instructions. or each library, 1µg of total RNA was used with the addition of 2µl of RNA Sequins spike-in mix A/B diluted 1:25 (Garvan Institute of Medical Research, Australia). Ribosomal RNA depleted RNA-seq libraries were generated using the KAPA RNA Hyperprep kit with RiboErase (Roche, cat. KK8560), following manufacturer's instructions. Each RNA-seq library was uniquely indexed using KAPA Dual-Indexed adapters (Roche, cat. KK8722). Libraries were quantified using a Qubit Fluorometer (Qubit dsDNA High Sensitivity kit) and fragment size was evaluated using an Agilent 2100 Bioanalyser (Agilent High Sensitivity DNA kit).
ENA-FIRST-PUBLIC
2019-05-01T04:02:51Z
ENA-LAST-UPDATE
2019-02-11T14:10:00Z
External Id
SAMEA5331852
INSDC center name
Bioinformatics Researcher
INSDC first public
2019-05-01T04:02:51Z
INSDC last update
2019-02-11T14:10:00Z
INSDC status
public
Submitter Id
E-MTAB-7655:ZFC4mut_1
broker name
ArrayExpress
cell line
ES cell line
cell type
mouse embryonic stem cell
common name
house mouse
developmental stage
embryo
genotype
ZFC4mut
sample name
E-MTAB-7655:ZFC4mut_1
scientific_name
Mus musculus