|
| Status |
Public on Dec 15, 2011 |
| Title |
PrefrontalCortex_Control_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
PrefrontalCortex_Wistar rat_replicate1
|
| Organism |
Rattus norvegicus |
| Characteristics |
gender: male
age: 10 week
brain region: Prefrontal cortex
strain: Wistar
average blood sugar [mmol/l]: 7.64
disease state: none
|
| Treatment protocol |
10 week old male Wistar rats (n=9) served as control animals in our experiments. 6 weeks old male Wistar rats were rendered diabetic via tail vein injection of STZ (n=9) (65 mg/kg iv, Sigma Chemical Co.) dissolved in 0.1M citrate buffer (pH 4.5)and served as models of type 1 diabetes mellitus. Only STZ-treated animals with plasma glucose concentrations above 15 mmol/L were considered diabetic and included in the study. 10 week old Goto Kakizaki rats (n=9) were used as a model of type 2 diabetes mellitus. Animals were anaesthetized by an intraperitoneal injection of pentobarbital sodium (5 mg /100 g body weight), then killed by decapitation.
|
| Growth protocol |
Experiments were performed on ten-week old male rats (weighing 286±60g). Rats were kept in a room with constant temperature (20°C), humidity and a 12-hour light-dark cycle, and were allowed to free access to standard chow and water.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Excised tissue samples were immediately fixed in RNA later RNA stabilization reagent (Qiagen, Valencia, CA). Total RNA was extracted from samples by homogenization using the RNeasy Kit (Qiagen, Valencia, CA), according to the manufacturer’s instructions. RNA integrity and purity were checked both by agarose gel electrophoresis and with an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). RNA was quantified using a NanoDrop-1000 spectrophotometer (Thermo scientific, Wilmington, DE). Samples of acceptable quality fulfilled the following criteria: OD260/280>1.8, OD260/230>1.8 and RIN>7.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 1000 ng RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions. Dye incorporation was controlled by a Nanodrop spectrophotometer; all samples were labeled with an efficiency of 10.2 – 17.5 pmol Cy3/μg cRNA.
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| |
|
| Hybridization protocol |
1650 ng of cRNA were hybridized to Agilent’s Rat Whole GenomeCustom Arrays for 17 hours at 65°C in a rotating Agilent hybridization oven.
|
| Scan protocol |
Hybridized arrays were imaged with Agilent’s Microarray Scanner in the extended dynamic range at 100% and 10% laser beam intensities at a resolutionof 5 μm.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters protocol GE1-v5_91_0806.
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| |
|
| Submission date |
Dec 14, 2011 |
| Last update date |
Dec 15, 2011 |
| Contact name |
Omar Abdul-Rahman |
| E-mail(s) |
abdulhun2000@yahoo.com
|
| Organization name |
Semmelweis University
|
| Department |
Department of Medical Chemistry, Molecular Biology and Pathobiochemistry
|
| Lab |
Sasvari lab
|
| Street address |
Tüzoltó utca 37-47
|
| City |
Budapest |
| ZIP/Postal code |
1094 |
| Country |
Hungary |
| |
|
| Platform ID |
GPL15011 |
| Series (1) |
| GSE34451 |
Altered gene expression profiles in the hippocampus and prefrontal cortex of type 2 diabetic rats |
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