The expression profiles of Drosophila Kc167 and SL2 cell lines were compared. Total RNA was extracted from dense cultures of Kc167 or SL2 cells which were maintained at 22C using Trizol Reagent (Invitrogen, Burlington, ON). Kc167 cells were grown in Schneider's Complete Drosophila Medium (Invitrogen) supplemented with 5% Fetal Bovine Serum (Invitrogen) and 20ug/mL Gentamicin (Sigma, Oakville, ON) whereas SL2 cells were grown in CCM3 media (HyClone, Logan, UT) supplemented with 20ug/mL Gentamicin. 80ug of RNA, quantified by A260 in an Ultrmark plate reader (Biorad, Mississauga, ON), was used in each direct labeling reaction (see http://www.flyarrays.com) to generate fluorescently labeled cDNA. The respective Cy3 (SL2) and Cy5 (Kc167) targets were suspended in 75 uL DIG EasyHyb solution (Roche, Mississauga, ON)and were competitively hybridized to the array for 16 hours in a CMT-GAPS hybridization chamber (Corning, Acton, MA). Scanning was performed with a ScanArray 4000 system (Perkin Elmer, Boston, MA) using the following settings: 10uM scan: Cy3(543 nm) 100-72, Cy5(633 nm) 95-60 (laser power-PMT Gain). Quantification was performed using QuantArray v3 software (Perkin Elmer) using the adaptive algorithm without background subtraction. The data in the file is not normalized. The ratio in the Value column was derived from normalized data (not shown) as discussed in the manuscript. Keywords = Kc167 Keywords = SL2 Keywords = Drosophila Lot batch = d7k2-070802
