Total RNA was extracted using TRI-reagent (Invitrogen)
Label
Cy3
Label protocol
Complementary RNA (cRNA) samples were linearly amplified and labeled with cyanine 3 starting from 100 ng of total RNA (One-Color Microarray-Based Gene Expression Analysis- Low Input Quick Amp Labeling” version 6.5, May 2010, Part Number G4140-90040 protocol)
Hybridization protocol
Following fragmentation, labeled cRNA were hybridized on Agilent “Mouse Gene Expression 8x60K Microarray” (Design ID: 028005), for 17 hrs, at 65°C under 10 rpm.
Scan protocol
Following washing, the slides were scanned using an Agilent G2565CA microarray Scanner System, at a 3 µm resolution in a 20-bit scan mode, according to the “AgilentG3_GX_1Color” protocol (scan resolution : 5 µm, Tiff : 20 bit).
Description
Gene expression in control
Data processing
Raw .tif images were then extracted using Agilent “Feature Extraction, version 10.10.1.1” following “GE1_1010_Sep10” protocol.
