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Nucleic Acids Research, 1990, Vol. 18, No. 3 501-505
© 1990

CHEMISTRY

The preparation of polyamide-oligonucleotide probes containing multiple non-radioactive labels

Jim Haralambidis*, Karin Angus, Scott Pownall, Lucy Duncan, Miao Chai and Geoffrey W. Tregear

Howard Florey Institute of Experimental Physiology and Medicine, University of Melbourne Parkville, Victoria 3052, Australia

* To whom correspondence should be addressed

Received October 24, 1989. Accepted November 27, 1989.

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).


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