3 protocols
Agilent Two-Color Microarray-Based Gene Expression (Quick Amp Labeling)
Version: 5.7
Agilent publication number: G4140-90050
URL: http://www.chem.agilent.com/Library/usermanuals/Public/G4140-90050_Two-Color_GE_5.7.pdf
This document describes Agilent's recommended procedures for the preparation and labeling of complex biological targets and hybridization, washing, scanning, and feature extraction of Agilent 60-mer oligonucleotide microarrays for microarray-based two-color gene expression analysis.
For each strain, total RNA was extracted from three pools of 30, three day old, non blood-fed females using Pico PureTM RNA Isolation Kit (Applied biosystems, Foster city, CA, USA). The strains were reared in parallel to minimize variation resulting from breeding conditions. Each biological replicate consisted of mosquitoes from distinct generations to control for stochastic variations. The quality and concentration of RNA was assessed using a 2100 Bioanalyzer (Agilent technologies, Santa Clara, CA, USA).
The VK and Ngousso strains were reared under standard conditions (25 + 2C, 70-80 % RH) in insectaries.