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Fast detection of a (CA) 18 microsatellite repeat in the IgE receptor gene by capillary electrophoresis with laser‐induced flurescence detection
Author(s) -
Klepárník Karel,
Malá Zdeňka,
Haváč Zdeněk,
Blažková Michaela,
Hollá Lydie,
Boček Petr
Publication year - 1998
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/elps.1150190218
Subject(s) - capillary electrophoresis , microbiology and biotechnology , microsatellite , laser induced fluorescence , gel electrophoresis , chromatography , agarose gel electrophoresis , chemistry , agarose , analytical chemistry (journal) , polymerase chain reaction , electrophoresis , fluorescence , dna , biology , gene , biochemistry , optics , allele , physics
Abstract The optimum separation conditions of polymerase chain reaction (PCR) products have been found for high‐speed capillary electrophoresis (CE) with laser‐induced fluorescence (LIF) detection. DNA fragments obtained after PCR amplification of the region covering the (CA) 18 microsatellite repeat in nitron 5 of the gene for FcERIβ, a high affinity glycoprotein receptor for IgE, located on chromosome 11 (11q13), were analyzed with the aim of investigating the repeat polymorphism. The results of polyacrylamide slab gel electrophoresis (PAGE), agarose gel electrophoresis, CE with absorbance detector and CE with LIF are compared. The CE with LIF proved to shorten analysis time by a factor of 100 when compared to slab gel electrophoresis. CE‐LIF utilizes a short capillary with an effective length of 6.3 cm and electric field strength from 100 to 550 V/cm. The respective PCR products of sizes from 116 to 210 base pairs (bp) were analyzed in 3 min.