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.
Scan protocol
Scan resolution 5um on an Agilent scanner.
Description
251643611654_S01_miRNA-v1_95_May07_2_3_BL Corresponds to GSM568274
Data processing
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.