Sample 1
Homo sapiens
Protocols: Cells were washed twice with cold PBS supplemented with 0.1% of EDTA (500 mM), harvested by scraping and collected in a tube. After a centrifugation at 3000 rpm for 5 min at 4°C, the nuclear fraction was extracted by resuspending the pellet in nuclear isolation Buffer (NIB) (1.28 M Sucrose, 40 mM Tris-HCl pH 7.5, 20 mM MgCl2 and 4% Triton X-100) supplemented with 100 U/ml RNAse inhibitor (RiboLock RNase Inhibitor, Invitrogen) and proteinase inhibitors (1mM PMSF and 1x PIC). Samples were incubated on ice for 20 min and then centrifuged at 2500 x g at 4°C for 15 min. Once removed the supernatant, nuclear pellets were resuspended in 400 µl of RIP Buffer (150 mM KCl, 25 mM Tris pH 7.4, 5 mM EDTA, 0.5 mM DTT e 0.5% NP40) supplemented with 100 U/ml RNAse and proteinase (1mM PMSF and 1x PIC) inhibitors. Cell nuclei were sonicated for 10 cycles (15\" ON and 15\" OFF) using Bioruptor (Diagenode, Denville, New Jersey, USA) and subsequently centrifuged at 13000 rpm for 10 min at 4°C. 1 ml of TRIzol (Life Technologies, Thermo Fisher) was added directly to the beads and RNA extraction was performed according to the manufacturer's guidelines TruSeq Stranded Total RNA Library Prep Gold (Cat. 20020599, Illumina, San Diego, California, USA)
ENA first public
2026-02-28
INSDC center name
University of Salerno
INSDC status
public
Submitter Id
E-MTAB-15354:Sample 1
broker name
ArrayExpress
cell line
MCF-7
cell type
mammary epithelial cell
collection date
not collected
common name
human
developmental stage
adult
disease
breast neoplasm
genotype
wild type genotype
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
breast
sample name
E-MTAB-15354:Sample 1
scientific_name
Homo sapiens