GeneChip® HT 3' IVT PLUS Reagent Kit

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Always keep thawed reagents and reaction tubes on ice

Upstream tips
Always keep thawed reagents and reaction tubes on ice

Publication protocol

The total RNA was extracted from the brain tissues previously collected using ISOGEN kit and quantified by Nanodrop 2000 spectrophotometer (Thermo Fisher scientific, Wilmington, NC, USA).
To elucidate the molecular mechanism underlying the effect of VEE on neuronal activities, we evaluated the total gene expression of brain tissues by performing microarray on RNAs previously extracted. The experiment was conducted according to the Affymetrix Genechip 3’ IVT PLUS reagent kit user’s guide. Briefly, the RNAs were reverse transcripted to generate double stranded DNA. The latter used as a template to synthesize the Biotin-labeled cRNA. After fragmentation of the labeled cRNA, the mixture was hybridized to the Affymetrix mouse 430 PM array strips (Affymetrix) for 16 h at 45 °C in the hybridization station. In Geneatlas Fluidics station, the hybridized arrays were washed and stained, then scanned using Geneatlas imaging station. The total number of genes analyzed by this method is 39,396 genes. All brain samples were analyzed by microarray. The data obtained were analyzed by Expression Console and Pathway Studio software and DAVID and Consensus Path databases.

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