2025-04-01
ENA first public
ERC000011
ENA-CHECKLIST
EMBL-EBI,
INSDC center name
public
INSDC status
ERS22946673
SRA accession
E-MTAB-14759:RS_R1_9
Submitter Id
ArrayExpress
broker name
A375
cell_line
melanocyte
cell_type
not collected
collection date
human
common name
Protocols: Cells were seeded at 0.5 million cells/well in 6-well plates and left overnight.  Before harvesting, cells were treated with Vemurafenib and Trametinib separately or in combination for 6 hours. Approximately 1 million cells were used as the starting material. Cells were washed twice with ice-cold PBS, scrapped with a cell scraper, and then centrifuged at 1000 × g for 3 min.  For RNA isolation, the RNeasy Mini Kit (Qiagen, Cat. No. 74104) was used per manufacturer instructions, along with QIAshredder columns for the homogenisation of cell lysates and DNase I treatment using DNase I kit. Following RNA extraction, the integrity of the isolated RNA was assessed using the Agilent 4200 TapeStation system according to the manufacturer's instructions, using 2 μL of a 1:150 dilution in H2O of each purified total RNA sample. The samples were tested further for cDNA synthesis using PrimeScript 1st strand cDNA synthesis kit (Takara Bio) followed by an RT-PCR on β-Actin using 2 μL of cDNA product.  mRNA-seq libraries were prepared from 500 ng of total RNA with the NEBNext RNA Ultra II kit coupled with the poly(A) mRNA Magnetic Isolation Module on the Beckman liquid handling robot i7 with these parameters: 10 min fragmentation and 10 PCR cycles.TBC
description
melanoma
disease
MED12 knockout
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
Homo sapiens
scientific_name
RS_R1_9
title