Total RNA was extracted from protoplasts in 1:1 (v/v) mixture of TE-saturated phenol and 0.1 M Tris-HCl buffer (pH 8.0) containing 0.1 M LiCl, 10 mM EDTA and 1 % SDS, and precipitated in 2.5 M LiCl according to Shirzadegan et al. (1991) except that extracted samples were not incubated in 80°C. Contaminated DNA was digested with DNase I (Takara, Otsu, Shiga, Japan). mRNA was purified from total RNA using Dynabeads Oligo (dT)25 (DYNAL, Dynal Biotech, Oslo, Norway) after DNase I treatment.
Label
biotin
Label protocol
0.3 microgram of mRNA was processed to produce biotinylated cRNA targets.
Hybridization protocol
Standard Affymetrix procedures
Scan protocol
Standard Affymetrix procedures
Description
Microarray analysis was performed to know how many gibberellin (GA)-responsive genes are inhibited by beta-Yariv reagent, a specific binder of plant arabinogalactan-proteins