Preparation of oligonucleotide microarrays on modified glass using a photoreactive heterobifunctional reagent, 1-N-(maleimidohexanoyl)-6-N-(anthraquinon-2-oyl) hexanediamine (MHAHD)

A novel approach for the construction of oligonucleotide microarrays under the influence of light and microwaves is described. For that purpose, a new heterobifunctional crosslinking reagent, 1-N-(maleimidohexanoyl)-6-N-(anthraquinon-2-oyl) hexanediamine (MHAHD), possessing a photoactive anthraquinone moiety on one terminus and an electrophilic maleimide group on the other one, was developed. The immobilization of oligonucleotides using the reagent, MHAHD, was realized via two routes (A and B). In order to speed up the immobilization procedure, the reaction between 3′- or 5′-mercaptoalkylated oligonucleotides and maleimide moiety of MHAHD was carried out under microwaves in just 15 min. The oligonucleotide arrays produced by both the routes were analyzed by hybridization experiments (hybridization efficiency 30.13%) and subsequently used for the discrimination of base mismatches. The constructed microarrays were found to possess good thermal stability (only ∼4.5% loss of fluorescence intensity observed after ten cycles).