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 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.
Klepárník
K
K
Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Brno. klep@ipm.cz
Malá
Z
Z
Havác
Z
Z
Blazková
M
M
Hollá
L
L
Bocek
P
P
D001483:Base Sequence; D004247:DNA; D004587:Electrophoresis, Agar Gel; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D005470:Fluorometry; D006801:Humans; D007834:Lasers; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D017455:Receptors, IgE / Q000235:genetics*; D013997:Time Factors
D017455:Receptors, IgE; D004247:DNA