Fast detection of a (CA)18 microsatellite repeat in the IgE receptor gene by capillary electrophoresis with laser-induced fluorescence detection

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 nitro...

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Veröffentlicht in:	Electrophoresis 1998-02, Vol.19 (2), p.249-255
Hauptverfasser:	Klepárník, K, Malá, Z, Havác, Z, Blazková, M, Hollá, L, Bocek, P
Format:	Artikel
Sprache:	eng
Schlagworte:	Base Sequence DNA Electrophoresis, Agar Gel Electrophoresis, Capillary - methods Electrophoresis, Polyacrylamide Gel Fluorescence Fluorometry Humans Lasers Microsatellite Repeats Molecular Sequence Data Receptors, IgE - genetics Time Factors
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creator	Klepárník, K Malá, Z Havác, Z Blazková, M Hollá, L Bocek, P
description	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 FcERIbeta, 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.
doi_str_mv	10.1002/elps.1150190218
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source	MEDLINE; Wiley Online Library Journals Frontfile Complete
subjects	Base Sequence DNA Electrophoresis, Agar Gel Electrophoresis, Capillary - methods Electrophoresis, Polyacrylamide Gel Fluorescence Fluorometry Humans Lasers Microsatellite Repeats Molecular Sequence Data Receptors, IgE - genetics Time Factors
title	Fast detection of a (CA)18 microsatellite repeat in the IgE receptor gene by capillary electrophoresis with laser-induced fluorescence detection
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