normalization data transformation protocol
Offset background subtracted, loess normalized. Data obtained from log2 of processed Ch1 signal / processed Ch2 signal. LIMMA software was used ID_REF = VALUE = Loess normalized log2 ratio (Ch1/Ch2)
array scanning protocol
Scanned on Chip Reader (Applied biosystems Laboratories) at 5 µm resolution
43°C, 16/20 h, hybridization buffer Northern Max (Ambion). Pre-hybridization for 1h in same conditions.
15 µg of total RNA were primed with 0.5 µg oligodT(19)VN primer at 70°C for 10 min, then reversed transcribed at 42°C for 2 h in the presence of 400 U ReverseAid MuLV H minus RTase (Fermentas), and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3-labeled dCTP or Cy5-label dCTP
nucleic acid extraction protocol
Total RNA extracted from the digestive gland of a single individual mussel using Trizol following manufacturer's instructions