Abstract
Oligonucleotide probes containing multiple non-radioactive labels have been prepared by utilising and extending the methods used to prepare polyamide-oligonucleotide conjugates. The probes were prepared by incorporating suitable amino acid residues, such as lysines, in the polyamide, which were then used as sites for the attachment of the non-radioactive labels. The procedures developed give control over the distance of the label from the oligonucleotide, and also the inter-label distance. The labels can be conveniently introduced while the substrate is still on the solid support. Even though fluorescent oligonucleotide probes prepared in this way carrying multiple carboxyfluorescein labels gave low levels of fluorescence due to quenching, the probes containing ten biotin labels gave a detection sensitivity of approximately 5 attomole (3 million molecules).
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Agrawal S., Christodoulou C., Gait M. J. Efficient methods for attaching non-radioactive labels to the 5' ends of synthetic oligodeoxyribonucleotides. Nucleic Acids Res. 1986 Aug 11;14(15):6227–6245. doi: 10.1093/nar/14.15.6227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Becker J. M., Wilchek M., Katchalski E. Irreversible inhibition of biotin transport in yeast by biotinyl-p-nitrophenyl ester. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2604–2607. doi: 10.1073/pnas.68.10.2604. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drinkwater C. C., Evans B. A., Richards R. I. Kallikreins, kinins and growth factor biosynthesis. Trends Biochem Sci. 1988 May;13(5):169–172. doi: 10.1016/0968-0004(88)90144-2. [DOI] [PubMed] [Google Scholar]
- Jablonski E., Moomaw E. W., Tullis R. H., Ruth J. L. Preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use as hybridization probes. Nucleic Acids Res. 1986 Aug 11;14(15):6115–6128. doi: 10.1093/nar/14.15.6115. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morawetz H. Studies of synthetic polymers by nonradiative energy transfer. Science. 1988 Apr 8;240(4849):172–176. doi: 10.1126/science.240.4849.172. [DOI] [PubMed] [Google Scholar]
- Urdea M. S., Warner B. D., Running J. A., Stempien M., Clyne J., Horn T. A comparison of non-radioisotopic hybridization assay methods using fluorescent, chemiluminescent and enzyme labeled synthetic oligodeoxyribonucleotide probes. Nucleic Acids Res. 1988 Jun 10;16(11):4937–4956. doi: 10.1093/nar/16.11.4937. [DOI] [PMC free article] [PubMed] [Google Scholar]