Flies from each vial (10 females) were homogenized with a FP-120 Fast Prep bead beater, according to manufacturer's protocols (Bio-101, Carlsbad, CA, USA), in 1.5 mL Trizol Reagent (Invitrogen, Carlsbad, CA, USA) and 150 mL chloroform. Double-stranded complementary DNA was prepared from 5 microgram total RNA using the SuperScript Choice System (Invitrogen), according to the manufacturer's instructions, except using an oligo-dT primer containing a T7 RNA polymerase promoter site.
Label
Biotin
Label protocol
Biotin-labeled complementary RNA (cRNA) was prepared using the BioArray High Yield RNA Transcript Labelling Kit (Enzo, Farmingdale, NY, USA). After the in vitro transcription (IVT) reaction, the unincorporated nucleotides were removed using RNeasy columns (Qiagen, Hilden, Germany).
Hybridization protocol
As described by Dyrskjot et al (2003), Nat. Gen. 33:90-96..
Scan protocol
As described by Dyrskjot et al (2003), Nat. Gen. 33: 90-96.
Description
Gene expression data from longevity selected flies
Data processing
raw data was GC-RMA normalized using the Bioconductor application for R
