|
| Status |
Public on Nov 17, 2015 |
| Title |
Veh 3 |
| Sample type |
RNA |
| |
|
| Source name |
rat hearts
|
| Organism |
Rattus norvegicus |
| Characteristics |
tissue: Heart
strain: Sprague-Dawley
gender: Male
treatment: exposed to vehicle
|
| Treatment protocol |
Diabetes mellitus was induced in Sprague-Dawley rats by a single injection of streptozotocin (STZ; 60 mg/kg) into the tail vein. The plasma glucose concentration in the rats was > 300 mg/dL and they were assigned to groups with diabetes mellitus and with DM and plasma glucose fluctuations induced by 24 hours of starvation and intraperitoneal insulin (0.5 IU/kg) injection.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from rat heart was extracted with a miRNeasy Mini kit (QIAGEN) following the manufacturer's instructions. RNA was quantified using a NanoDrop-1000 spectrophotometer.
|
| Label |
Cy3
|
| Label protocol |
One hundred ng of total RNA was labeled with Cy3 using a miRNA labeling and hybridization kit (Agilent Technologies) according to the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
After Cy3-labeling, RNA was dried up and re-solved with 18 ul RNase-free water. And then, 4.5 ul of 10x Gene expression blocking agent and 22.5 ul of 2x hybridization buffer HI-RPM were added to the RNA sample, and the mixture hybridized to Agilent rat miRNA Microarrays (Grid ID19159, Rat Rel. 10.1, Agilent Technologies) for 20 hours at 55°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 5 minute at room temperature with GE Wash Buffer 1 (Agilent Technologies) and 5 minute with 37°C GE Wash buffer 2 (Agilent Technologies).
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565BA) using one color scan setting for 8x15k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to 5% and 100%, extended dynamic range scan mode).
|
| Description |
miRNA expression in the heart of control rat
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.5.3.1 (Agilent Technologies) using default parameters (protocol miRNA-v1_95_May07 and Grid: 019159_D_20081121) to obtain background subtracted Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Sep 11, 2012 |
| Last update date |
Nov 17, 2015 |
| Contact name |
Yasushi Teshima |
| Organization name |
Oita University
|
| Street address |
1-1 Idaigaoka, Yufu
|
| City |
Oita |
| ZIP/Postal code |
8700026 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10344 |
| Series (1) |
| GSE40779 |
The expression profiles of miRNAs in the heart with glucose fluctuation compared with sustained hyperglycemia. |
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