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Status |
Public on Sep 01, 2017 |
Title |
Heart Ctrl no treatment sample 2 (mouse n° 217) |
Sample type |
RNA |
|
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Channel 1 |
Source name |
Heart Ctrl no treatment
|
Organism |
Mus musculus |
Characteristics |
Sex: male genotype: Ctrl (non-transgenic) treatment: no treatment treatment duration: 7 days tissue: Heart
|
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.
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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 |
tissues: 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.
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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 |
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.
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Submission date |
Oct 17, 2011 |
Last update date |
Sep 01, 2017 |
Contact name |
Smail Messaoudi |
E-mail(s) |
smail.messaoudi@gmail.com
|
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 (2) |
GSE33022 |
Cardiac aldosterone and corticosterone regulated genes in mice with cardiomyocytes targeted mineralocorticoid receptor overexpression (MR-Cardio mice) and their controls (Ctrl mice) |
GSE34049 |
Cardiac aldosterone regulated genes in mice with cardiomyocytes targeted mineralocorticoid receptor overexpression (MR-Cardio mice) and their controls (Ctrl mice) |
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