Sample 2
Homo sapiens
Protocols: A375-P cells treated with either 1 uM PLX4720 or 0.01% DMSO for 36 h. Cells were washed once with PBS, briefly incubated with trypsin at 37°C, re-suspended in ice-cold 2.5% FCS/PBS supplemented with either 1 uM PLX4720 or 0.01% DMSO and transferred into a 15 ml Falcon tube. Cells were counted, pelleted at 400 g for 4' at 4°C. Samples were filtered through a round-bottom test tube with a cell strainer snap cap (BD Falcon) to ensure single cell suspension. Sorts were performed on a BD FACSAriaIII or a BD FACSAriaIIIu Cell Sorter (BD Biosciences) using a 100 um nozzle. 200'000 cells were sorted directly into 2 ml Eppendorf tubes containing 700 L RA1 (lysis buffer) from the NucleoSpin® RNA II RNA isolation kit (Macherey-Nagel) containing 70 l TECP (Sigma-Aldrich). Eppendorfs were vortexed regularly to ensure cell lysis. RNA was isolated according to the manufacturer's protocol. The integrity of the RNA samples was checked using the Bioanalyzer system. Their concentration was measured using Quant-IT RiboGreen RNA Assays (Life Technologies). Libraries for Illumina sequencing have been prepared with 200 ng total RNA input material using the TruSeq Stranded Total RNA Library Prep kit Ribozero (Illumina) and quality checked using the Fragment Analyzer standard sensitivity NGS kit (AATI).
ENA-FIRST-PUBLIC
2019-08-08T04:04:57Z
ENA-LAST-UPDATE
2018-12-03T11:14:42Z
External Id
SAMEA5145604
INSDC center name
Institute of Molecular Health Sciences, ETH Zurich
INSDC first public
2019-08-08T04:04:57Z
INSDC last update
2018-12-03T11:14:42Z
INSDC status
public
Submitter Id
E-MTAB-7453:Sample 2
broker name
ArrayExpress
cell line
A375-P
cell type
epithelial cell
common name
human
disease
melanoma
organism part
skin
sample name
E-MTAB-7453:Sample 2
scientific_name
Homo sapiens