title: Oligonucleotides site-specifically spin-labeled at 5'-terminal or internucleotide linkage and their use in gene analyses.
abstract: Spin-labeled oligonucleotides (S-probes) were synthesized and examined as DNA probes to monitor hybrid formation. TEMPO was introduced either at the internucleotide linkage of 5'-terminus (Type 1) or at the 5'-terminal hydroxyl group (Type 2) and both types of S-probes were used in this study. The presence of target DNA was detected in solution by EPR spectroscopy for both types of S-probes. Hybridization of the S-probes resulted in notable broadening of EPR line width, accompanied by a decrease in the EPR signal height ratio for I(-1)/I(0).I(-1)/I(0) of S-probes having no spacer between oligonucleotide and TEMPO decreased more markedly than that of S-probes with a spacer, indicating that TEMPO should be introduced to an oligonucleotide directly to monitor hybrid formation. When M13mp8 single-stranded DNA with or without an EcoRI recognition site was selected as a target DNA, hybrid formation was detected only for DNA containing EcoRI site in solution using spin-labeled oligonucleotides.
authors.affiliation: Department of Polymer Science and Engineering, Kyoto Institute of Technology, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Murakami
authors.forename: M
authors.initials: M
authors.lastname: Mukae
authors.forename: S
authors.initials: S
authors.lastname: Nagahara
authors.forename: Y
authors.initials: Y
authors.lastname: Konishi
authors.forename: H
authors.initials: H
authors.lastname: Ide
authors.forename: K
authors.initials: K
authors.lastname: Makino
mesh_terms: D017104:Bacteriophage M13; D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis; D004578:Electron Spin Resonance Spectroscopy; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry*; D012996:Solutions; D013113:Spin Labels
chemical_list: D004279:DNA, Viral; D015345:Oligonucleotide Probes; D012996:Solutions; D013113:Spin Labels