|
| Status |
Public on Dec 01, 2016 |
| Title |
WT |
| Sample type |
RNA |
| |
|
| Source name |
WT_dorsal skin
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain background: Wistar Bonn/Kobori (WBN/Kob)
genotype/variation: wild type
gender: male
age: 25wk
tissue: dorsal skin
|
| Treatment protocol |
Drosal skins were prepared from WT and Ht rats at 25 weeks of age, frozen immediately, and stored at −80 °C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs of skin samples were extracted using TRIzol reagent (Thermo Fisher Scientific, Rockford, IL, USA), purified using RNeasy Mini kit (Qiagen, Hilden, Germany) following the manufacturer's instructions. Extracted RNA was quantified using a NanoDrop spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 100 ng RNA using Low Input Quick Amp Labeling Kit, one-color (Agilent) according to the manufacturer's instructions. Dye incorporation and cRNA yield were checked with the NanoDrop Spectrophotometer.
|
| |
|
| Hybridization protocol |
0.6 ug of Cy3-labelled cRNA was fragmented at 60°C for 30 minutes, hybridized to Agilent Whole Rat Genome Oligo Microarrays (SurePrint G3 Rat GE 8x60K v2 Microarray) at 65°C for 17 hour in rotating at 10 rpm using Gene Expression Hybidization Kit (Agilent) following the manufacturers instructions. After hybridization, microarrays were washed using Gene Expression Wash Buffers Pack (Agilent) following manufacturers instructions.
|
| Scan protocol |
Slides were scanned immediately after washing on the SureScan microarray scanner (G2600D) using one color scan setting.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.5.1.1 (Agilent) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Jul 12, 2016 |
| Last update date |
Dec 01, 2016 |
| Contact name |
Masato Wakabayashi |
| E-mail(s) |
masato.wakabayashi@shionogi.co.jp
|
| Organization name |
Shionogi & Co., Ltd.
|
| Street address |
3-1-1, Futaba-cho
|
| City |
Toyonaka-shi |
| State/province |
Osaka |
| ZIP/Postal code |
561-0825 |
| Country |
Japan |
| |
|
| Platform ID |
GPL22145 |
| Series (1) |
| GSE84294 |
Decreases in 15-lipoxigenase metabolites in Olmsted syndrome model rats |
|
