|
| Status |
Public on Dec 16, 2014 |
| Title |
35D_1 |
| Sample type |
RNA |
| |
|
| Source name |
15 berries incubated for 10 days under 35ºC/Dark condition
|
| Organism |
Vitis labrusca x Vitis vinifera |
| Characteristics |
cultivar: Pione
tissue: grape berry just beginning to show color
|
| Treatment protocol |
Grape berries just beginning to show color were collected from mature vines of 'Pione'; The berries were incubated for 10 days under one of four conditiond; 15ºC/Light, 15ºC/Dark, 35ºC/Light, 35ºC/Dark.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from frozen samples by the hot borate method (Wan and Wilkins, 1994).
|
| Label |
Cy3
|
| Label protocol |
Total RNA (250 ng) was labelled with Cy3 according to the instructions for the Quick Amp Labeling Kit (Agilent, Palo Alto, CA). Labeled cRNA was purified using the RNeasy Mini Kit (Qiagen, Hilden, Germany).
|
| |
|
| Hybridization protocol |
Glass slides were hybridized overnight at 65 °C in a hybridization buffer containing a portion of Cy3 -labeled cRNA. After hybridization, slides were washed in Agilent Gene Expression Wash Buffer 1 (Agilent) with 0.005% Triton X-102 for 1 min at room temperature and Agilent Gene Expression Wash Buffer 2 (Agilent) with 0.005% Triton X-102 for 1 min at 37 °C.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565A) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.5 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Dec 15, 2014 |
| Last update date |
Dec 16, 2014 |
| Contact name |
Hiroshi Fujii |
| E-mail(s) |
hfujii@affrc.go.jp
|
| Organization name |
Institute of Fruit Tree Science, NARO
|
| Department |
Division of Citrus Research
|
| Street address |
485-6 Okitsunakacho, Shimizu
|
| City |
Shizuoka |
| State/province |
Shizuoka |
| ZIP/Postal code |
424-0292 |
| Country |
Japan |
| |
|
| Platform ID |
GPL19557 |
| Series (1) |
| GSE64153 |
Microarray Analysis for the Screening of Genes Inducible by Light or Low Temperature in Post-veraison Grape Berries |
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