Klepárnik K, Malá Z, Pribyla L, Blazková M, Vasků A, Bocek P
Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Brno.
Electrophoresis. 2000 Jan;21(1):238-46. doi: 10.1002/(SICI)1522-2683(20000101)21:1<238::AID-ELPS238>3.0.CO;2-E.
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 600C 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.
开发了短毛细管快速变性电泳的方法和仪器，并以内皮素1基因短串联重复多态性的检测为例进行了说明。在有效长度为2.5 cm的毛细管中，于60℃温度和600 V/cm电场强度下，42秒内实现了检测二核苷酸重复多态性所需的两个核苷酸的分辨率。因此，与传统平板凝胶电泳相比，在具有激光诱导荧光检测的短毛细管中使用变性电泳可将分析时间缩短200倍。所开发的方法和仪器有利于在临床诊断和遗传群体筛查中应用，在这些领域中，适用于自动化的快速分析仪器至关重要。
