LEJ1-overexpressed Arabidopsis Flowers with anther stage from 1 to 12
Growth protocol
unopened flower buds of 5-week-old wild-type grown under LD conditions at 24?.
Extracted molecule
total RNA
Extraction protocol
Trizol extraction of total RNA was performed according to the manufacturer's instructions. And then purified with the RNeasy Mini kit.
Label
biotin
Label protocol
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 microg total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
Hybridization protocol
Following fragmentation, 15 microg of cRNA were hybridized for 16 hr at 45? on Arabidopsis ATH1 Genome Array. After hybridization, the arrays were washed in a GeneChip Fluidics Station 450 with a non-stringent wash buffer at 25? followed by a stringent wash buffer at 50?. After washing, the arrays were stained with a streptavidin-phycoerythrin complex. After staining, intensities were determined with a GeneChip scanner, controlled by GCOS Affymetrix software.
Scan protocol
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
Description
Gene expression data from unopened flowers of wild-type plant
Data processing
The data were analyzed with Affymetrix GeneChip Operating Software. For normalization, data from each expression array were scaled, so that the overall fluorescence intensity across each chip was equivalent (average target intensity set at 500). The One-Sided Wilcoxon's Signed Rank test was employed to generate the Detection p-value.
