Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S.

Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S. Research Abstract Details 

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Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S. Abstract Text:

Cameron Mroske,John Muci,Jicheng Wang,Kai Li,Wenjia Song,Jin Yan,Jinong Feng,Qiang Liu,Steve S Sommer,

Although DOVAM-S (detection of virtually all mutations-SSCP) in effect detects all mutations and is less costly than direct sequencing, the technique currently requires the use of radioactivity. F-DOVAM-S (fluorescent DOVAM-S) was developed to replace the isotopic label with fluorescence and to increase throughput via dye color multiplexing. As proof of principle, two multitemperature slab gel electrophoresis conditions were evaluated through the blinded analysis of mutations in the factor IX (FIX) genes of 88 hemophilia B (HB) patients and 7 wild-type controls. Using only two conditions, it was determined that F-DOVAM-S had a detection sensitivity of 97%. It is anticipated that when three or four optimized conditions are employed, F-DOVAM-S will detect all mutations. Three patient samples were multiplexed per well using three different fluorescent dyes (6FAM, VIC, and NED), demonstrating that it is possible to analyze up to 44 kb of diploid, color-coded amplification product per gel lane. This value corresponds to a throughput of approximately 4 Mb of DNA analyzed per 96-well gel, which is approximately triple that of conventional radiolabeled DOVAM-S. Throughput is further enhanced by the rapidity at which the fluorescent signal can be captured and the resultant multicolor chromatograms analyzed. Given these data, F-DOVAM-S has the potential to be a particularly powerful technology for clinical diagnosis because it allows the mutation analysis of multiple patients to be performed within 24h.

Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S. Publishing Authors By Initials

C Mroske,J Muci,J Wang,K Li,W Song,J Yan,J Feng,Q Liu,SS Sommer,

For similar investigative techniques: epidemiologic methods: statistics as topic: sensitivity and specificity research abstracts see: investigative techniques: epidemiologic methods: statistics as topic: sensitivity and specificity research

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Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Analytical biochemistry

VOLUME: 368

Page Numbers: 250-7

Journal Abbreviation: Anal. Biochem.

ISSN: 0003-2697

DAY: 26

MONTH: 05

YEAR: 2007

Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S. Information

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LANGUAGE: eng

NlmUniqueID: 370535

Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S. Keywords Mesh Terms:

KEYWORDS: Sensitivity and Specificity

MESH TERMS: metabolism

Chemical & Substance for Abstract: Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S. Information

Substance Name: Factor IX

Registry Number: 9001-28-9

Grant and Affiliation Information for Toward a fluorescent single-strand conformation polymorphism technique that detects all mutations: F-DOVAM-S.

AFFILIATION: Department of Molecular Genetics and Department of Molecular Diagnosis, City of Hope National Medical Center, Duarte, CA 91010, USA.

Country: United States

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MEDLINETA: Anal Biochem

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