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DNA-DNA Hybridization in Real Time Using BIAcore

✍ Scribed by S.J. Wood

Publisher
Elsevier Science
Year
1993
Tongue
English
Weight
352 KB
Volume
47
Category
Article
ISSN
0026-265X

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✦ Synopsis

Sheila J. Wood
ERDEC Research Directorate, SCBRD-RTE, Aberdeen Proving Ground, Maryland 21010 Received February 13, 1992; accepted May 12, 1992

Hybridization between complementary DNA oligonucleotides was performed in the real time monitoring flow analysis system, BIAcore. Avidin was covalently immobilized to a dextran matrix and served as the reaction surface. Biotinylated oligonucleotides, introduced within a fluid stream, were captured by the immobilized avidin. Complementary DNA oligonucleotides successfully hybridized with the biotinylated base probes. The hybridization reaction was carried out at room temperature and positive signals were obtained within (7 \mathrm{~min}). In addition, biotinylated and complementary DNA oligonucleotides were prehybridized prior to injection. Hybrid pairs were captured by the avidin surface giving results commensurate with the concentrations injected. Control experiments showed that identically sized, but dissimilar DNA oligonucleotides did not hybridize with the biotinylated base probe. Nonspecific attachment of DNA directly to the reaction surface was not observed. This is the first account of specific hybridization occurring in real time within a fluid flow system detectable in less than (10 \mathrm{~min}) at room temperature. 1993 Academic Press, Inc.

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