Protocols: Cells were harvested from respective cell cultures. Human fibroblasts were cultured in Minimum Essential Medium Eagle (MEM; Gibco) supplemented with 15 % fetal bovine serum (FBS; Gibco), 2 mM L-glutamine (Gibco), and the antibiotics penicillin (100 units/ml) and streptomycin (100 μg/ml) (Pen/Strep; Gibco) at 37°C. Human wild type osteosarcoma cells (U2OS) were cultured in Dulbecco's Modified Eagle's Medium-high glucose (DMEM-high glucose; Sigma) supplemented with 10 % FBS and the antibiotics Pen/Strep at 37°C. BRAT1 KO gene edited U2OS cell lines were prepared using Cas9 and guide sequences designed in CRISPR direct (https://crispr.dbcls.jp). Total RNA was isolated from 1×106 cells by miRNeasy Micro Kit (Qiagen) with an additional DNase I treatment, according to the manufacturer's instructions. Sequencing libraries were prepared with the SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian user manual (Takara Bio USA, Inc).
ENA-FIRST-PUBLIC
2022-07-27
ENA-LAST-UPDATE
2022-07-27
External Id
SAMEA9424341
INSDC center alias
Laboratory of Genomics and Bioinformatics Institute of Molecular Genetics of the Czech Academy of Sciences
INSDC center name
Laboratory of Genomics and Bioinformatics Institute of Molecular Genetics of the Czech Academy of Sciences
INSDC first public
2022-07-27T12:58:55Z
INSDC last update
2022-07-27T12:58:55Z
INSDC status
public
Submitter Id
E-MTAB-10750:WT #1
broker name
ArrayExpress
cell_line
U-2 OS
cell_type
osteosarcoma cell
common name
human
developmental stage
adult
disease
osteosarcoma
familial relationship
none
genotype
wild type genotype
isolate
not applicable
organism part
bone
sample name
E-MTAB-10750:WT #1
sex
female