Total RNA was extracted by Trizol reagent (Invitrogen, Carlsbad, CA, USA).
Label
Cy3
Label protocol
100ng of total RNA were dephosphorylated, denaturalizated, treated with ligase T4 and labelled with Cy3 for 2 hours at 16ºC.
Hybridization protocol
Hybridization settings: 20 hours, at 55ºC, at 20rpm. Blocking agent, hybridization and washing buffers was performed according to Agilent's recommendations.
Limma package was used to import the data. After between-array median normalization by the R program, significant analysis of microarray (SAM) Excel macro was used.
