E-MTAB-15334:Cpt_Cas_rep1
2025-09-05
ENA first public
ERC000011
ENA-CHECKLIST
Laboratory of Molecular Medicine and Genomics, Department of Medicine, Surgery and Dentistry ‘Scuola Medica Salernitana’, University of Salerno, Baronissi (SA), Italy
INSDC center name
public
INSDC status
ERS25309177
SRA accession
E-MTAB-15334:Cpt_Cas_rep1
Submitter Id
ArrayExpress
broker name
HeLa
cell line
uterine smooth muscle cell
cell type
not collected
collection date
human
common name
Protocols: Cervix adenocarcinoma cell line (HeLa), human triple-negative breast cancer (TNBC) model cell line (MDA-MB-231) and pleural effusion of hormone-dependent human breast carcinoma (MCF-7) were obtained the American Type Culture Collection (ATCC). HeLa and MBA-MD 231 were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Thermo Fisher Scientific, Waltham, MA, USA). MCF7 cell line was cultured in Dulbecco&#39;s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA). All media were supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific), 100 U/mL penicillin, 100 µg/mL streptomycin, and 2 mM L-glutamine. Cells were maintained at 37 °C in a humidified incubator with 5% CO₂. Total RNA was extracted from frozen cell samples using Qiagen RNeasy Plus Universal mini kit following manufacturer’s instructions (Qiagen, Hilden, Germany).     RNA samples were quantified using Qubit 3.0 Fluorometer (ThermoFisher Scientific, Waltham, MA, USA) and RNA integrity was checked with 4200 TapeStation (Agilent Technologies, Palo Alto, CA, USA). The RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina using manufacturer’s instructions (New England Biolabs, Ipswich, MA, USA). Briefly, mRNAs were initially enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3’ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles. The sequencing library was validated on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 3.0 Fluorometer (ThermoFisher Scientific, Waltham, MA, USA) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA).
description
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
uterine cervix
organism part
Homo sapiens
scientific_name
CHK1 inhibitor+Castalin
stimulus
Cpt_Cas_rep1
title