VFB mutant analysis

Transcript profiling analysis of vfb (Vier F-Box) mutant seedlings compared to wild type using Arabidopsis ATH1 GeneChip array. Keywords: 10 day old light grown seedlings, wild type and mutant Three replicate samples were harvested and prepared from 10 day-old light grown wild type and vfb mutant seedlings. RNA for micro array analysis was extracted using the RNeasyTM kit (Quiagen, Hilden, Germany). Complementary RNA was prepared from 6 µg of total RNA as described in Affimetrix _Expression Analysis Technical Manual using the One-Cycle Target Labeling and Control Reagents (Affimetrix, Santa Clara, CA). In brief, double-stranded cDNA was synthesized. Biotin-labeled target cRNA was prepared by cDNA in vitro transcription in the presence of biotinylated UTP and CTP. After purification, cRNA was fragmented and used to hybridize to the Arabidopsis ATH1 GeneChip array (Affimetrix, Santa Clara, CA). Hybridization, washing, staining, scanning and data collection were performed in an Affymetrix GeneChip® Fluidics Station 450 and GeneArray® Scanner. The microarray computaional analysis was performed on *.CEL and *.CHP data files