|
| Status |
Public on Dec 20, 2007 |
| Title |
Endosperm 16DAF; Seed development; Rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
Barley Endosperm 16DAF
|
| Organism |
Hordeum vulgare |
| Characteristics |
Genotype: Barke cultivar
Age: 16DAF
|
| Treatment protocol |
From the early stages of developing caryopsis, the maternal tissues, i.e. mostly pericarp (p), and the filial endosperm/embryo fraction as embryo sac (es) were separated by manual dissection as described by Sreenivasulu et al. (2002). At later stages 16 and 25 DAF endosperm+aleurone (e/a) and embryo+scutellum (em) fractions were prepared.
|
| Growth protocol |
Barley plants were cultivated in growth chambers at 20oC/18°C under a 16 h light/8 h dark cycle until seed set. The developmental stage of a caryopsis was determined from the mid-region of the ear and assigned as 4, 8, 16 and 25 days after flowering (DAF).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions, followed by Rneasy spincolumn cleanup
|
| Label |
Probe synthesis, labelling and hybridisation were performed according to manufacturer’s protocols (Affymetrix, Santa Clara, CA) at the UCI MicroArray Core Facility, Irvine, Ca.]
|
| Label protocol |
The BioArray High-Yield RNA Transcript Labeling Kit (Enzo Diagnostics) was then used to synthesize biotin-labeled cRNA from template cDNA by in vitro transcription.
|
| |
|
| Hybridization protocol |
10 µg labeled, fragmented cRNA was then hybridized at 45°C with rotation for 16 hours in an Affymetrix GeneChip Hybridization Oven 320 on Affymetrix Barley1 Genechip arrays. The arrays were washed and stained using streptavidin phycoerythrin on an Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000
|
| Description |
Gene expression data from barley endosperm 16 DAF
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
Oct 18, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Björn Usadel |
| E-mail(s) |
usadel@mpimp-golm.mpg.de
|
| Organization name |
Max Planck Institute of Molecular Plant Physiology
|
| Street address |
Am Mühlenberg 1
|
| City |
Potsdam |
| ZIP/Postal code |
14476 |
| Country |
Germany |
| |
|
| Platform ID |
GPL1340 |
| Series (1) |
| GSE9365 |
Expression data from barley maturing and germinating grains |
|
