title: A01225
organism: Homo sapiens
description: Protocols: Serums from patients belong to the Azure Trial (BIG 01/04) were analysed 13. For the aim of this study, 48 serums were selected based on patient's outcome by primary tumour resection for a 10-years-period. The serum has been aliquoted in 1mL tubes and stored at -80°C in Sheffield under approved ethical conditions until assayed. The absence of serum hemolysis and fibrin was assessed by macroscopic visual analysis. Total RNA was extracted from 200 µl serum using the miRCURY Biofluids RNA Isolation Kit (Exiqon®, Vedbaek, Denmark) according to the manufacturer's recommendations. For each serum sample, the miRNome analysis was performed by Next Generation Sequencing (NGS) using 20 µl of the 45 µl total RNA extracted from 200 µl serum by the miRCURY Biofluids RNA Isolation Kit. The NGS libraries were constructed and sequenced at the ProfileXpert platform (www.profilexpert.fr). Total RNA (20 l) was converted into miRNA NGS libraries using the QIAseq miRNA library kit (QIAGEN). Adapters containing UMI (Unique Molecular Index) were ligated to the 3'-OH and 5'-PO4 RNA prior to the reverse-transcriptase reaction to generate cDNA. The cDNA was amplified with a 20 cycle PCR with primers containing sample specific indexes allowing the library identification in the sequencing analysis. After the cDNA clean-up on QMN beads and the amplification step the size range of the libraries was analyzed on the Bioanalyser 2100 (Agilent) to perform quality control (QC). Samples were then quantified by specific qPCR using a standard curve (0.002-20 nM) and the libraries were pooled in equimolar concentrations (2nM of each library). Quality control of the library synthesis was validated as the library size of 173 pb without primer dimers at 150 pb and with a minimal amount of 1 nM.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-06-10
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-06-10
attributes.tag: External Id
attributes.value: SAMEA9147862
attributes.tag: INSDC center alias
attributes.value: Postdoctoral Research Associate
attributes.tag: INSDC center name
attributes.value: Postdoctoral Research Associate
attributes.tag: INSDC first public
attributes.value: 2023-06-10T00:17:27Z
attributes.tag: INSDC last update
attributes.value: 2023-06-10T00:17:27Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10555:A01225
attributes.tag: age
attributes.value: 52.9
attributes.tag: allocation
attributes.value: Standard treatment alone
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cohort
attributes.value: One
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: Not Available
attributes.tag: disease
attributes.value: Breast Cancer
attributes.tag: disease recurrence
attributes.value: No first Recurrence
attributes.tag: estrogen-receptor positive breast cancer
attributes.value: ER positive
attributes.tag: individual
attributes.value: Not Available
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: Blood
attributes.tag: sample name
attributes.value: E-MTAB-10555:A01225
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: female
attributes.tag: tumor grading
attributes.value: 2
attributes.tag: tumor stage
attributes.value: T1