DESIGN AND SYNTHESIS OF POLYACRYLAMIDE-BASED OLIGONUCLEOTIDE SUPPORTSFOR USE IN NUCLEIC-ACID DIAGNOSTICS

Polyacrylamide supports, in a range of pore sizes, were investigated a s nucleic acid affinity matrices for the detection of target DNA or RN A sequences using a sandwich hybridization format. Bromoacetyl and thi ol oligonucleotide derivatives were covalently linked to sulfhydryl- a nd bromoacetyl-polyacrylamide supports with greater than 95% end-attac hment efficiencies. These polyacrylamide-oligonucleotide supports were further derivatized with anionic residues to provide multi-functional supports which show low non-specific binding for non-complementary nu cleic acids. While all the polyacrylamide-oligonucleotide supports cap ture complementary oligonucleotides with high affinity, the pore size was found to be a critical parameter in sandwich hybridization reactio ns. The superior hybridization characteristics of the Trisacryl suppor t was ascribed to a combination of its macroporous nature, hydrophilic ity and the terminal attachment of its capture oligonucleotides.