Bridged Oligonucleotides with Smoothed Hybridization Properties as a Tool for Analysis of Nucleotide Sequences

Resumo

Bridged oligonucleotides that contain nonnucleotide inserts, i.e., diethylene glycol phosphate residues (DEG insert) have been studied. These oligonucleotide derivatives were shown to smooth the melting temperatures of their DNA duplexes of various nucleotide composition. The DEG insert has been shown to have almost no effect on the sequence specificity of modified oligonucleotides and regulate only their hybridization properties. It has been demonstrated that bridged oligonucleotides immobilized on microparticles can be successfully used to specifically and efficiently reveal DNA templates during the parallel solid-phase hybridization analysis. The use of bridged oligonucleotides along with DNA-dependent enzymes (Taq DNA polymerase and T4 DNA ligase) in this analysis has led to pronounced discrimination of wrong DNA molecules due to the covalent attachment of the label to the carrier, thus increasing the reliability of the hybridization analysis.