-transparent.png){kind=logo}
A series of oligonucleotides containing biotin-11-dUMP at various positions were synthesized and compared in quantitative, colorimetric hybridization-detection studies. A deoxyuridine phosphoramidite containing a protected allylamino sidearm was synthesized and used in standard, automated synthesis cycles to prepare oligonucleotides with allylamino residues at various positions within a standard 17-base sequence. Biotin substituents were subsequently attached to the allylamino sidearms by reaction with N-biotinyl-6-aminocaproic acid N-hydroxysuccinimide ester. These oligomers were hybridized to target DNA immobilized on microtiter wells (ELISA plates), and were detected with a streptavidin-biotinylated horseradish peroxidase complex using hydrogen peroxide as substrate and o-phenylenediamine as chromogen. We found that the sensitivity of detection of target DNA by biotin-labeled oligonucleotide probes was strongly dependent upon the position of the biotin label. Oligonucleotides containing biotin labels near or off the ends of the hybridizing sequence were more effective probes than oligonucleotides containing internal biotin labels. An additive effect of increasing numbers of biotin-dUMP residues was found for some labeling configurations.
A F Cook, and
E Vuocolo, and
C L Brakel
January 1991,
Nucleic acids symposium series,
A F Cook, and
E Vuocolo, and
C L Brakel
January 1989,
Molekuliarnaia biologiia,
A F Cook, and
E Vuocolo, and
C L Brakel
January 1989,
Acta histochemica. Supplementband,
A F Cook, and
E Vuocolo, and
C L Brakel
August 1997,
Nucleic acids research,
A F Cook, and
E Vuocolo, and
C L Brakel
June 2001,
BioTechniques,
A F Cook, and
E Vuocolo, and
C L Brakel
August 1990,
Nucleic acids research,
A F Cook, and
E Vuocolo, and
C L Brakel
September 2020,
Organic & biomolecular chemistry,
A F Cook, and
E Vuocolo, and
C L Brakel
July 1999,
Bioorganic & medicinal chemistry,
A F Cook, and
E Vuocolo, and
C L Brakel
August 1995,
BioTechniques,
A F Cook, and
E Vuocolo, and
C L Brakel
June 1984,
DNA (Mary Ann Liebert, Inc.),
