|
| Status |
Public on Sep 08, 2016 |
| Title |
Heart_Ctrl + aldosterone treatment_ sample 1 (mouse n° 203) |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Heart_Ctrl + aldosterone treatment
|
| Organism |
Mus musculus |
| Characteristics |
gender: male
treatment: Aldosterone
treatment duration: 7 days
tissue: Heart ventricle
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared from ventricle samples with the RNeasy Mini Kit (Qiagen S.A) according to the manufacturer's instructions and its quality was assessed with Bioanalyzer 2100 (Agilent technologies)
|
| Label |
Cy5
|
| Label protocol |
Labelling of the RNAs weas performed according to Agilent's instruction. Briefly, 100 ng of total RNA + spikes were primed with 1.8µl of T7 promoter primer mix at 70°C for 10 min, then reversed transcribed at 40°C for 2 h in presence of a mix containing first strand buffer, 20 mM DTT, 2 mM DNTP mix and AffinityScript rnase block mix. Reaction was stopped by heating samples at 70° foc 15 min. New synthetized cDNA was then labelled and amplified at 40° for 2h in presence of a mix containing transcription buffer, NTP mix, 20mM DTT, T7 RNA polymerase and CY3-CTP or CY5-CTP.
|
| |
|
| Channel 2 |
| Source name |
Total RNA pooled from several tissues (liver, brain, heart kidney, spleen), labeled with Cyanine-3, used as reference
|
| Organism |
Mus musculus |
| Characteristics |
tissue composition of reference: liver, brain, heart kidney, spleen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared from ventricle samples with the RNeasy Mini Kit (Qiagen S.A) according to the manufacturer's instructions and its quality was assessed with Bioanalyzer 2100 (Agilent technologies)
|
| Label |
Cy3
|
| Label protocol |
Labelling of the RNAs weas performed according to Agilent's instruction. Briefly, 100 ng of total RNA + spikes were primed with 1.8µl of T7 promoter primer mix at 70°C for 10 min, then reversed transcribed at 40°C for 2 h in presence of a mix containing first strand buffer, 20 mM DTT, 2 mM DNTP mix and AffinityScript rnase block mix. Reaction was stopped by heating samples at 70° foc 15 min. New synthetized cDNA was then labelled and amplified at 40° for 2h in presence of a mix containing transcription buffer, NTP mix, 20mM DTT, T7 RNA polymerase and CY3-CTP or CY5-CTP.
|
| |
|
| |
| Hybridization protocol |
825 ng of Cy5-labelled samples and 825 ng of Cy3-labelled references were incubated at 60° for 30 min in fragmentation buffer. Hybridization buffer was then added, and sample/reference couples (1 couple per microarray) were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequential
|
| Scan protocol |
Scanned on an G2505 Agilent scanner.
Images were quantified using Agilent Feature Extraction Software (version v 10.7.3.1).
|
| Description |
Sample 6
Hearts were rinsed in cold PBS, frozen in liquid nitrogen, and kept at –80°C.
|
| Data processing |
Agilent Feature Extraction Software (v 10.7.3.1) was used for background subtraction and LOWESS normalization.Data were processed using genespring software v11.0.
|
| |
|
| Submission date |
Sep 09, 2013 |
| Last update date |
Sep 08, 2016 |
| Contact name |
Smail Messaoudi |
| E-mail(s) |
[email protected]
|
| Phone |
+1 613 562 5800
|
| Organization name |
University of Ottawa
|
| Department |
BMI
|
| Street address |
451 smyth road
|
| City |
Ottawa |
| State/province |
Ontario |
| ZIP/Postal code |
K1H 8M5 |
| Country |
Canada |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE50709 |
Cardiac aldosterone regulated genes in mice |
|
