|
| Status |
Public on Jan 12, 2010 |
| Title |
Cardiac_RV_WT_Rep3 |
| Sample type |
RNA |
| |
|
| Source name |
cardiac right ventricle free wall
|
| Organism |
Mus musculus |
| Characteristics |
tissue: cardiac right ventricle free wall
strain: 129P2/OlaHsd
age: 12 weeks
gender: Male
transgene: wild-type
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from tissue samples from mice aged 10 to 12 weeks, using the Nucleospin RNA II kit according to manufacturer’s instructions (Clontech). RNA quality was assessed using the Agilent 2100 Bioanalyzer, and RNA concentration was measured by spectrophotometry (Nanodrop).
|
| Label |
Digoxigenin-UTP
|
| Label protocol |
Digoxigenin-UTP labeled cRNA was generated from 1 microg of total RNA using Applied Biosystems Chemiluminescent RT-IVT labeling kit according to manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Each micro-array was first prehybridized at 55C for 1 hr in hybridization buffer with blocking reagent, after which 13 microg of labeled cRNA was hybridized to each array in a 1.5mL volume at 55C for 16 hr. After hybridization, the arrays were washed with hybridization wash buffer and chemiluminescence rinse buffer.
|
| Scan protocol |
Chemiluminescence detection, image acquisition and analysis were performed using Applied Biosystems Chemiluminescence Detection Kit and Applied Biosystems 1700 Chemiluminescent Microarray Analyzer
following manufacturer’s protocol.
|
| Description |
Single-color chemiluminescent signals were quantified, corrected for background and spatially normalized. Applied Biosystems Expression System software was used to extract assay signal, and assay signal to noise ratio values from the micro-array images. Bad spots flagged by the software were removed from the analysis.
|
| Data processing |
The raw signal of the set of genes was log transformed and quantile-normalized across arrays, and genes were further filtered by standard expression array signal to noise threshold (S/N greater than 3 in at least one sample).
|
| |
|
| Submission date |
Jan 04, 2010 |
| Last update date |
Jan 11, 2010 |
| Contact name |
Brendon Scicluna |
| E-mail(s) |
brendon.scicluna@um.edu.mt
|
| Organization name |
University of Malta
|
| Street address |
Msida campus
|
| City |
Msida |
| ZIP/Postal code |
MSD2020 |
| Country |
Malta |
| |
|
| Platform ID |
GPL2995 |
| Series (1) |
| GSE19741 |
Genetic background effect on Scn5a mutant cardiac right ventricle gene expression |
|
