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Ultrafast detection of microsatellite repeat polymorphism in endothelin 1 gene by electrophoresis in short capillaries
Author(s) -
Klepárník Karel,
Malá Zdeňka,
Přibyla Lubomír,
Blažková Michaela,
Vašků Anna,
Boček Petr
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000101)21:1<238::aid-elps238>3.0.co;2-e
Subject(s) - capillary electrophoresis , microsatellite , microbiology and biotechnology , biology , electrophoresis , chromatography , gel electrophoresis , population , high resolution melt , single nucleotide polymorphism , polymerase chain reaction , gene , chemistry , genetics , genotype , allele , medicine , environmental health
The methodology and instrumentation for fast denaturing electrophoresis in short capillaries was developed and exemplified by detection of short tandem repeat polymorphism in the endothelin 1 gene. The resolution of two nucleotides, which is required for the detection of a dinucleotide repeat polymorphism, was achieved in a capillary of an effective length of 2.5 cm at a temperature of 60°C and an electric field strength of 600 V/cm in 42 s. Thus, the use of denaturing electrophoresis in short capillaries with laser‐induced fluorescence detection resulted in a reduction of analysis time by a factor of 200 when compared to the conventional slab gel electrophoresis. The developed methodology and instrumentation is advantageous for an implementation in clinical diagnostics and genetic population screening where fast analytical instrumentation amenable to automation is of paramount importance.