Design and synthesis of polyacrylamide-based oligonucleotide supports for use in nucleic acid diagnostics

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Abstract
Polyacrylamlde supports, in a range of pore sizes, were Investigated as nucleic acid affinity matrices for the detection of target DNA or RNA sequences using a sandwich hybridization format. Bromoacetyl and thiol oligonucleotide derivatives were covalently linked to sulfhydryl- and bromoacetyl-polyacrylamide supports with greater than 95% end-attachment efficiencies. These polyacrylamide-oligonucleotide supports were further derivatlzed with anionlc residues to provide multi-functional supports which show low non-specific binding for non-complementary nucleic acids. While all the polyacrylamide-oligonucleotide supports capture complementary ollgonucleotides with high affinity, the pore size was found to be a critical parameter In sandwich hybridization reactions. The superior hybridization characteristics of the Trisacryl support was ascribed to a combination of Its macroporous nature, hydrophiliclty and the terminal attachment of Its capture ollgonucleotides.