BioProject

Analysis of gene expression in Arabidopsis thaliana leaf, stem and flower tissues. More...

Analysis of gene expression in Arabidopsis thaliana leaf, stem and flower tissues. Columbia (Col-0) Arabidopsis thaliana plants were grown at a density of 4 plants per 5 inch square pot either in a growth chamber or a green house set to 25*C by day, 20*C by night. Days were set to a 16hr photoperiod with 125 micro mol m-2s-1 fluorescent irradiation. Expanding leaves were harvested 15 days post germination in the middle of the photoperiod (3 replicates). Expanding upper 2" of the stem with siliques and pedicels removed were harvested 29 days post germination in the middle of the photoperiod (4 replicates). Developed flowers and unopened buds were harvested 29 days post germination in the middle of the photoperiod (4 replicates). RNA and Microarray Methods: Total RNA was extracted from the plants using the Trizol method (Invitrogen, Ramonell et al. (2002) Mol. Plant Pathol. 3: 301) and purified with a silica membrane column (Qiagen, RNeasy). Twenty micrograms biotinylated complementary RNA (cRNA) was prepared as described (Hernan et al. (2003) Cancer Res. 63, 140) from the purified total RNA. The resulting cRNA was used to hybridize ATH1 Arabidopsis GeneChips (Affymetrix) using the manufacturer's protocols. The array images were analyzed with the Affymetrix Microarray Suite 5.0 software with the target intensity set to 500. Normalized signal intensities were generated by multiplying signal intensities by ratio figure which set GapC (ID_REF = 258588_s_at) to 8500. Keywords: other Less...