Flies were entrained in LD (light: dark) condition for 3-4 days and harvested at two time points: ZT3 and ZT15.
Extracted molecule
total RNA
Extraction protocol
Total RNA was prepared (30 heads per sample) using Trizol reagent (Invitrogen) according to the manufacturer's protocol.
Label
biotin
Label protocol
RNA was then used to prepare a probe that was hybridized to Drosophila 2.0 gene expression arrays (Affymetrix) according to the manufacturer's protocol.
Hybridization protocol
standard Affymetrix protocol
Scan protocol
standard Affymetrix protocol
Data processing
CEL files were imported to Affymetrix Expression Console program, and data were normalized using Robust Multichips Analysis. Differential expression was tested for probes that had expression levels above 4.0 (log2 scale) in at least one group (mutants or WT). All other probes were excluded from the analysis.
