Klepárník K, Malá Z, Havác Z, Blazková M, Hollá L, Bocek P
Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Brno.
Electrophoresis. 1998 Feb;19(2):249-55. doi: 10.1002/elps.1150190218.
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.
已找到用于聚合酶链反应（PCR）产物的最佳分离条件，该条件适用于配备激光诱导荧光（LIF）检测的高速毛细管电泳（CE）。对位于11号染色体（11q13）上的FcERIβ（一种高亲和力IgE糖蛋白受体）基因第5外显子中覆盖（CA）18微卫星重复序列区域进行PCR扩增后获得的DNA片段进行了分析，目的是研究重复序列多态性。比较了聚丙烯酰胺平板凝胶电泳（PAGE）、琼脂糖凝胶电泳、配备吸光度检测器的CE和配备LIF的CE的结果。与平板凝胶电泳相比，配备LIF的CE分析时间缩短了100倍。CE-LIF使用有效长度为6.3 cm的短毛细管，电场强度为100至550 V/cm。在3分钟内分析了大小从116至210个碱基对（bp）的各个PCR产物。
