TETHERED DYE GENE PROBES

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Authors
  1. Piunno, P.A.E.
  2. Krull, U.J.
  3. Lee, W.
Corporate Authors
Defence Research Establishment Suffield, Ralston ALTA (CAN);Toronto Univ, Toronto ONT (CAN) Dept of Chemistry
Abstract
The research presented in this report concentrates on the synthesis of transduction agents for conversion of selective nucleic acid hybridization events into measurable analytical signals. The use of transduction elements as opposed to lables provides for a diagnostic assay method which is internally calibrated and obviates the need for washing or other reagent treatments protocols. The two transduction elements focused on were polyether tethered analogues of the fluorescent intercalant double-stranded DNA ligands ethidium bromide and YOYO-1. Proof of concept for this strategy has been provided by a hydrocarbon tethered analogue of ethidium bromide attached to thymidylic acid icosanucleotides (Deoxyribothymidylic Acid Icosanucleotide) immobilized on fused silica optical fibers in an optical biosensor configuration. Transduction of hybridization events betwen the immobilized Deoxyribothymidylic Acid Icosanucleotide and complementary sequences was done in real time with full analytical signal generation completed within a time of ca.6 minutes. Synthetic protocols developed towards the synthesis of polyether tethered phosphoramidite analogues of these fluorophores is herein disclosed. In this way, oligonucleotide probes functionalized with polyether tethered fluorescent transduction elements for use in hybridization assays may readily be fabricated by use of an automated DNA synthesizer within time periods of hours.
Keywords
Transduction agents;Diagnostic assay methods;Polyether tethered analogues;Biosensors;DNA synthesizers
Report Number
DRES-CR-97-27 — Contractor Report
Date of publication
01 May 1997
Number of Pages
71
DSTKIM No
98-00449
CANDIS No
507097
Format(s):
Hardcopy;Document Image stored on Optical Disk

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