Tissue source: dissected adult thoracicoabdominal ganglia (the fused thoracic ganglia that are easily dissected from the ventral surface of the thorax after removing the head).
Canton-S flies were reared at 22oC on a 12:12hr light:dark regime on standard Drosophila diet, 1 week after adult emergence. Tissues were isolated by hand dissection from equal numbers of male and female flies (except in the case of gonads). Multiple samples were pooled to obtain enough total RNA.
Total RNA was extracted by RNEasy Micro kit (Invitrogen). 2ug of total RNA was used for labeled cRNA synthesis, according to Affymetrix protocol.
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45[o]C on GeneChip Drosophila Genome 2 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450. GeneChips were scanned using the GeneChip scanner 3000 7G.