|
| Status |
Public on Dec 03, 2012 |
| Title |
LeafBlade_58DAT_18:00 |
| Sample type |
RNA |
| |
|
| Source name |
Leaf blade, 58 days after transplanting, 18:00
|
| Organism |
Oryza sativa Japonica Group |
| Characteristics |
cultivar: Nipponbare
organ/tissue: Leaf Blade
growth/development stage: 58 days after transplanting
sampling time: 18:00
|
| Growth protocol |
All plants were grown in paddy field in Tsukuba, Japan (~36o N latitude) during a normal rice growing season in 2008.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using RNeasy Plant Mini Kit (QIAGEN) according to the manufacturer's protocol, quantitied using NanoDrop ND-1000 UV-VIS spectrophotometer (NanoDrop) and quality-checked using Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA).
|
| Label |
Cy3
|
| Label protocol |
400 ng of total RNA was added with one-color spike-mix and labeling was performed using Quick Amp Labeling Kit, One-Color (Agilent) in the presence of cyanine-3-CTP accoring to the manufacturer's protocol. The cyanine-3-labeled cRNA was purified by RNeasy Mini Kit (QIAGEN), and quantified using NanoDrop ND-1000 UV-VIS spectrophotometer.
|
| |
|
| Hybridization protocol |
For hybridization, 1650 ng of cyanine 3-labeled cRNA is fragmented and hybridized on the slide of rice 4x44K microarray RAP-DB (G2519F#15241) at 65°C for 17 hours using the Agilent Gene Expression Hybridization Kit.
|
| Scan protocol |
Slides were scanned on an Agilent G2505B DNA microarray scanner using one-color scan setting for 4x44K array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green PMT: XDR Hi 100%; XDR Lo 10%).
|
| Description |
Gene expression of leaf blade at 58 days after transplanting, 18:00
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.5.3.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended processed signal intensities. The processed signal intensities were used for 75 percentile normalization and log2 transformation with GenespringGX11(Agilent).
|
| |
|
| Submission date |
Feb 23, 2012 |
| Last update date |
Dec 03, 2012 |
| Contact name |
Yutaka Sato |
| E-mail(s) |
satoyu@affrc.go.jp
|
| Organization name |
National Institute of Agrobiological Sciences
|
| Street address |
2-1-2 Kannondai
|
| City |
Tsukuba |
| State/province |
Ibaraki |
| ZIP/Postal code |
305-8602 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6864 |
| Series (1) |
| GSE36040 |
Diurnal and circadian gene expression profile of leaf throughout the entire growth of rice in the field |
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