|
| Status |
Public on Oct 09, 2014 |
| Title |
AAA- SMC AAA - 3 |
| Sample type |
RNA |
| |
|
| Source name |
Patient T1
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Abdominal aortic aneurysm
cell type: smooth mucle cells
|
| Treatment protocol |
Human abdominal aortic aneurysm biopsy samples were harvested during open surgery treatment. Human abdominal non aneurysmal aortas were harvested during multiretrieval organs, with the written consent of their families and the authorization of the French Biomedicine Agency (PFS 11-004).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from 10 mm2 of LCM-isolated areas with 200 μL TRI Reagent® (Ambion), according to the manufacturer’s instructions. RNA quality (RNA integrity number, RIN) and quantity were systematically checked for each sample with Pico 6000 Chips and then Small RNA Chips on the BioAnalyzer 1200 (Agilent Technologies) and using the NanoDrop 2000 (Thermo Scientific), respectively and according to the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
The miRNA Complete Labeling Kit (Agilent Technologies) required 100 ng of total RNA per sample. After dephosphorylation, they were labeled with Cyanine 3-pCp at 16°C for 2 hours, with a T4 ligase. Micro Bio-Spin 6 chromatography columns (Bio-Rad Laboratories, Hercules, CA) were used to purify the labeled RNA.
|
| |
|
| Hybridization protocol |
The miRNA Complete Hybridization Kit (Agilent Technologies) required 100 ng of total RNA per sample. Purified labeled miRNAs were prepared with 10 × Blocking Agent and 2 × Hi-RPM hybridization buffer and incubated with the array in hybridization chambers (Agilent) for 20 hours at 55 °C.
|
| Scan protocol |
SureScan microarray scanner (Agilent Technologies).
|
| Description |
Microdissected areas
|
| Data processing |
Raw data were acquired with Feature Extraction Software (Version 10.7.3.1). Calculation of the total gene signal for each miRNA replicated probe subtracted background signals. We used the R package script AgiMicroRna for normalization.
|
| |
|
| Submission date |
Oct 08, 2014 |
| Last update date |
Oct 09, 2014 |
| Contact name |
David Hot |
| E-mail(s) |
david.hot@pasteur-lille.fr
|
| Organization name |
Institut Pasteur de Lille
|
| Department |
Center for Infection and Immunity of Lille
|
| Lab |
Transcriptomics and Applied Genomics
|
| Street address |
1 rue du professeur Calmette
|
| City |
Lille |
| ZIP/Postal code |
59000 |
| Country |
France |
| |
|
| Platform ID |
GPL14943 |
| Series (1) |
| GSE62179 |
MicroRNA profiling of cells isolated from human abdominal aortic aneurysms. Let-7f, a new potential circulating biomarker |
|
