This document describes Agilent's dual-mode Gene Expression Microarray platform and the method for gene expression analysis using RNA isolated from human whole blood. The platform combines the ease and simplicity of one-color experimental design with the capability of two-color sensitivity for detection of small gene expression changes. Agilent's platform now includes a method for the analysis of differential gene expression from human whole blood, which is compatible with the PAXgene™ Blood RNA System. The PAXgene Blood RNA System consists of a blood collection tube and nucleic acid purification kit for the collection, storage, and transport of blood and stabilization of intracellular RNA for subsequent isolation and purification. Combined with the PAXgene System, Agilent's amplification and labeling technology, in conjunction with the high sensitivity and specificity of 60-mer oligonucleotide microarrays, offer a total solution for gene expression in whole blood without the need for additional steps or kits to remove globin transcripts. Titration experiments using Agilent RNA spike-in control targets demonstrate the sensitivity of detection.
This document details a method for analyzing differential gene expression from human whole blood using Agilent's dual-mode Gene Expression Microarray platform. The approach is compatible with the PAXgene Blood RNA System to stabilize RNA and includes a workflow that eliminates the need for additional steps to remove globin transcripts. The note demonstrates that the sensitivity for detecting low-abundance transcripts in whole blood RNA compares favorably with other cell and tissue sources.
The procedure requires collecting 2.5 ml of whole blood into a PAXgene Blood RNA Tube.
Typical RNA yields from 2.5 ml of whole blood range from 4 to 15 pg.
The ratio of absorbance at 260 nm and 280 nm was consistently 1.9 - 2.0.