| High-throughput genotyping of factor V Leiden mutation by ultrathin-layer agarose gel electrophoresis. | |
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MedLine Citation:
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PMID: 10486761 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Ultrathin-layer agarose gel electrophoresis is a novel combination of the established methodologies of slab gel electrophoresis and capillary gel electrophoresis. This new format provides a multilane separation platform with rapid analysis time and excellent sensitivity by using laser-induced fluorescence scanning detection system. Sample injection onto the ultrathin-layer separation platform is easily accomplished by membrane mediated loading technology. In this paper, we demonstrate the sensitivity and high-throughput fashion of this novel separation and detection system for rapid genotyping of the coagulation factor V Leiden mutation by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis. The PCR amplified fragment from exon 10 of the factor V gene was digested by the Mnl I restriction enzyme, followed by automated ultrathin-layer agarose gel electrophoresis analysis with "in migratio" fluorescent labeling during the separation process. Due to its speed and automation, this method should be considered for large scale screening of factor V Leiden mutation. |
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Authors:
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T Lengyel; M Sasvari-Szekely; A Guttman |
Publication Detail:
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Type: Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Journal of chromatography. A Volume: 853 ISSN: 0021-9673 ISO Abbreviation: J Chromatogr A Publication Date: 1999 Aug |
Date Detail:
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Created Date: 1999-10-26 Completed Date: 1999-10-26 Revised Date: 2009-01-15 |
Medline Journal Info:
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Nlm Unique ID: 9318488 Medline TA: J Chromatogr A Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 519-25 Citation Subset: IM |
Affiliation:
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Institute of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis Medical School, Budapest, Hungary. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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DNA Mutational Analysis
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methods* Electrophoresis, Agar Gel / methods* Electrophoresis, Polyacrylamide Gel / methods Factor V / analysis, genetics* Fluorescence Genotype Lasers Polymerase Chain Reaction Polymorphism, Restriction Fragment Length |
| Chemical | |
Reg. No./Substance:
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0/factor V Leiden; 9001-24-5/Factor V |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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