Use of hybridization kinetics for differentiating specific from non-specific binding to oligonucleotide microarrays

Abstract
Hybridization kinetics were found to be significantly different for specific and non‐specific binding of labeled cRNA to surface‐bound oligonucleotides on microarrays. We show direct evidence that in a complex sample specific binding takes longer to reach hybridization equilibrium than the non‐ specific binding. We find that this property can be used to estimate and to correct for the hybridization contributed by non‐specific binding. Useful applications are illustrated including the selection of superior oligonucleotides, and the reduction of false positives in exon identification.