characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-09-01T00:13:45Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-09-01T00:13:45Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA10110056
characteristics.characteristic: External Id
characteristics.text: INSERM U1242 OSS (Oncogenesis, Stress and Signaling) - Universite de Rennes1
characteristics.characteristic: INSDC center name
characteristics.text: 2022-09-01T00:13:45Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-09-01T00:13:45Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS7766614
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-10940:miR-Control
characteristics.characteristic: Submitter Id
characteristics.text: 49
characteristics.unit: year
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: MP41
characteristics.characteristic: cell line
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: MP41 cell line was obtained from Decaudin’s lab at Curie Institute, Paris, France. Mel202 and 92.1 cell line were obtained from European Collection of Authenticated Cell Cultures (ECACC) (Merck). All cell lines were maintained in humidified air (37°C, 5% CO2). Uveal melanoma cell lines were maintained in RPMI-1640 medium (GibcoTM, ThermoFisher, MA) supplemented with 20% Fetal Bovine Serum (FBS) (EurobioScientific) and 1% Penicillin-Streptomycin (PS) antibiotics (GibcoTM, ThermoFisher). All cell lines have been routinely tested for mycoplasma contamination (Mycoplasma contamination detection kit; rep-pt1; InvivoGen). Experiments were performed on MP41 cells according to the protocol published by Judy Lieberman’s lab (Tan SM, Lieberman J. Capture and identification of mirna targets by biotin pulldown and RNA-seq. In: Methods in Molecular Biology. 2016. p. 211-28.) but with minor modifications. 15 millions of MP41 cells were seeded in 3 X 150mm dishes 5 millions each) and were transfected the next day with 100nM of biotinylated miR-16 or miR-CTR (Dharmacon). Next, they were harvested ~24h post transfection. Cells from the 3 dishes were treated separately. Meanwhile, magnetic beads (Streptavidine Dynabeads™ M-280 DYNAL™, ThermoFisher Scientific) were washed and blocked according to the protocol. Cell lysate and washed beads were incubated for 4 hours at 4°C on a rotating agitator. All next steps: the precipitation and the purification of coupled RNA was performed according to the protocol. We pooled the 3 same conditions (from the 3 transfected plates) in the end of the RNA purification. Purified RNA was quantified using quantified using a NanoDrop 1000 Spectrophotometer (ThermoFisher Scientific) Experiments were performed on MP41 cells according to the protocol published by Judy Lieberman’s lab (Tan SM, Lieberman J. Capture and identification of mirna targets by biotin pulldown and RNA-seq. In: Methods in Molecular Biology. 2016. p. 211-28.).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: uveal melanoma
characteristics.characteristic: disease
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: eye
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: miR-Control
characteristics.characteristic: title
characteristics.text: biotinylated miR-Control transfection
characteristics.characteristic: treatment