. Total RNA was isolated using RNeasy Mini Kit (Qiagen).
Label
Cy5
Label protocol
Five mg of total RNA was used for Cy5-labeled aRNA preparation per manufacturer instruction (Ambion MessageAmp™ aRNA Amplification Kit, Ambion Inc., Austn, TX, USA).
The arrays were then scanned using GenePix 4000B (Molecular Devices, Sunnyvale, CA, USA) and the fluorescent intensities are extracted from the generated images by following the instructions and the conditions described in HOA User Guide. The raw intensity data were input to GeneSpring GX Version 7.3.1 (Agilent, Foster City, CA, USA).
Description
H48-0603004525 580 33 460 10 10_0635
Data processing
The raw intensity data were input to GeneSpring GX Version 7.3.1 (Agilent, Foster City, CA, USA). The averaged intensities for each probe then were normalized by quantile normalization. Differentially expressed genes were identified and filtered based on t-tests (p< 0.05) and fold-change cutoffs of 1.7 fold.
