069 Covalent binding of DNA to CovaLink NH microwell plates ? methods and applications

Abstract

Hybridization methods can be separated into two groups: liquid phase and solid phase hybridization. Solid phase hybridization is the most popular method due to easy handling. However, solid phase hybridization has one major disadvantage, i.e. the nucleic acid molecules are bound non-covalently with several attachment sites between the solid phase and the nucleic acid molecules. Multiple attachment sites are supposed to affect the hybridization capability negatively. Ideally, the most suitable binding between the nucleic acid molecules and the solid phase is a covalent bond between the solid phase and the nucleic acid molecules. Preferably, the nucleic acid molecules should be bound at the Y-end or the Y-end. Binding of DNA molecules at the Y-end has been described. The formation of a phosphoramidate bond between the amines and the Y-end is possible by carbodiimide condensation [1]. This technique has been used more or less successfully to bind DNA to various solid phases [ 2 5]. Recently, we have demonstrated that DNA molecules are covalently bound to CovaLink NH microwell plates by carbodiimide condensation [6]. Binding of 1.2 picomoles oligonucleotide (30b) per well was obtained. Binding of DNA to microwell plates in a specific way adds an interesting aspect to the solid phases suitable for hybridiza-