Do-it-yourself: construction of a custom cDNA macroarray platform with high sensitivity and linear range

摘要

BackgroundResearch involving gene expression profiling and clinical applications, such as diagnostics and prognostics, often require a DNA array platform that is flexibly customisable and cost-effective, but at the same time is highly sensitive and capable of accurately and reproducibly quantifying the transcriptional expression of a vast number of genes over the whole transcriptome dynamic range using low amounts of RNA sample. Hereto, a set of easy-to-implement practical optimisations to the design of cDNA-based nylon macroarrays as well as sample ³³P-labeling, hybridisation protocols and phosphor screen image processing were analysed for macroarray performance.