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Status |
Public on Nov 30, 2011 |
Title |
Liver Nrf2 knock-out High Fat Diet, replicate 2 |
Sample type |
RNA |
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Source name |
Liver Nrf2 knock-out 6 months High Fat Diet, replicate 2
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Organism |
Mus musculus |
Characteristics |
treatment: high fat diet tissue: liver genotype: Nrf2 KO gender: male genetic background: C57BL/6J
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Treatment protocol |
Liver from wild type and Nrf2-KO mice was excised after perfusion with ice-cold saline and then it was immersed in RNA later solution (Ambion,Foster City, CA).
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Growth protocol |
Mice were housed in the animal facility in the University of Patras Medical School in temperature-, light- and humidity-controlled rooms with a 12-h light/dark cycle.
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated using a guanidinium thiocyanate based method using the Trizol reagent (Invitrogen, Carlsbad,CA) and was further purified using the Rneasy minikit (Qiagen,Hilden,Germany) following the manufacturer's instructions.
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Label |
Cy3
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Label protocol |
Labelling of total RNA was performed in Atlas Biolabs (Berlin, Germany) using the following kit: Low Input Quick Amp Labeling Kit, One Color (#5190-2305), Agilent Technologies, using 100 ng of total RNA as starting material, according to the manufacturer´s instructions (Manual One-Color Microarray-Based Gene Expression Analysis, Low Input Quick Amp Labeling, Version 6.5, Mai 2010, Agilent Technologies). cRNA synthesis was regarded successful provided that =1.65 μg of cRNA with a Cy3-incorporation rate =8.0 pmol/μg cRNA were synthesized.
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Hybridization protocol |
Hybridization was performed in Atlas Biolabs (Berlin,Germany) as follows;1.65 μg of Cy3-labelled cRNA were fragmented according to the manufacturer´s instructions, and 1.425 μg of fragmented cRNA were hybridized (conc: 15 ng/μl). Hybridization conditions: 65°C, 10 rpm, 17 h; in an Agilent hybridization oven
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Scan protocol |
Scanning was performed using an Agilent DNA Microarray Scanner (Model G2505C) and Agilent´s Scan Control Software (version A.8.3.1) in Atlas Biolabs (Berlin,Germany).
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Description |
Gene expression after 6 months on high-fat diet
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Data processing |
Primary data analysis was performed using Agilent´s Feature Extraction Software (version 10.7.3.1) in Atlas Biolabs (Berlin,Germany).
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Submission date |
Nov 09, 2011 |
Last update date |
Nov 30, 2011 |
Contact name |
Dionysios Chartoumpekis |
E-mail(s) |
dchart@upatras.gr
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Organization name |
Lausanne University Hospital CHUV
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Department |
Service of Endocrinology Diabetes and Metabolism
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Street address |
Ave de la Sallaz 8
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City |
Lausanne |
ZIP/Postal code |
1011 |
Country |
Switzerland |
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Platform ID |
GPL11202 |
Series (1) |
GSE33575 |
Hepatic gene expression profiles after long-term high-fat diet feeding in wild type and Nrf2 knock-out mice. |
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