Beyond Affymetrix arrays: expanding the set of known hybridization isotherms and observing pre-wash signal intensities.

ABSTRACT: Microarray hybridization studies have attributed the nonlinearity of hybridization isotherms to probe saturation and post-hybridization washing. Both processes are thought to distort 'true' target abundance because immobilized probes are saturated with excess target and stringent washing removes loosely bound targets. Yet the paucity of studies aimed at understanding hybridization and dissociation makes it difficult to align physicochemical theory to microarray results. To fill the void, we first examined hybridization isotherms generated on different microarray platforms using a ribosomal RNA target and then investigated hybridization signals at equilibrium and after stringent wash. Hybridization signal at equilibrium was achieved by treating the microarray with isopropanol, which prevent

SUBMITTER: Pozhitkov AE 

PROVIDER: S-EPMC2836560 | biostudies-literature | 2010 Mar

REPOSITORIES: biostudies-literature