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PMID: 2594760 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 23 ·1989-12-00 ·Pages 9178-82

Chehab FF, Kan YW

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.

MeSH Terms
Base Sequence Chromosome Deletion DNA/blood,genetics,isolation & purification DNA, Viral/genetics Female Genetic Complementation Test Globins/genetics Humans Hydrops Fetalis/blood,genetics Leukocytes/analysis Molecular Sequence Data Nucleic Acid Amplification Techniques Oligonucleotide Probes Pregnancy Spectrometry, Fluorescence/methods
Chemicals
DNA, Viral Oligonucleotide Probes Globins DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Chehab F F
Applied Biosystems, Research and Development, Foster City, CA 94404.
Kan Y W
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35 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.23.9178
Published
1989-12-00
Pages
9178-82
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC298457
Subset
IM
Grants
NIDDK NIH HHS · DK16666 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.23.9178 PMC Full Text View journal details
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