Detection of specific DNA sequences by fluorescence amplification: a color complementation assay.

Abstract:

:We have developed a color complementation assay that allows rapid screening of specific genomic DNA sequences. It is based on the simultaneous amplification of two or more DNA segments with fluorescent oligonucleotide primers such that the generation of a color, or combination of colors, can be visualized and used for diagnosis. Color complementation assay obviates the need for gel electrophoresis and has been applied to the detection of a large and small gene deletion, a chromosomal translocation, an infectious agent, and a single-base substitution. DNA amplification with fluorescent oligonucleotide primers has also been used to multiplex and discriminate five different amplified DNA loci simultaneously. Each primer set is conjugated to a different dye, and the fluorescence of each dye respective to its amplified DNA locus is scored on a fluorometer. This method is valuable for DNA diagnostics of genetic, acquired, and infectious diseases, as well as in DNA forensics. It also lends itself to complete automation.

authors

Chehab FF,Kan YW

doi

10.1073/pnas.86.23.9178

subject

Has Abstract

pub_date

1989-12-01 00:00:00

pages

9178-82

issue

23

eissn

0027-8424

issn

1091-6490

journal_volume

86

pub_type

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