Butler J M, McCord B R, Jung J M, Lee J A, Budowle B, Allen R O
Department of Chemistry, University of Virginia, Charlottesville, USA.
Electrophoresis. 1995 Jun;16(6):974-80. doi: 10.1002/elps.11501601163.
Capillary electrophoresis (CE) is an analytical technique which provides rapid, high resolution analysis of amplified DNA fragments produced by the polymerase chain reaction (PCR). In this study, two internal standards are used as size markers to bracket und precisely size PCR products. The technique is applied to typing PCR products from the short tandem repeat locus HUMTH01. HUMTH01 consists of five to seven major alleles in the size range of 179-203 bp, with each allele four bp apart. Using this genetic marker, a population containing 97 individuals was examined with both polyacrylamide gel electrophoresis and CE. Identical genotypes were obtained with both techniques demonstrating the reliability of CE in DNA typing applications. The DNA analysis took place in sets of 10 with a calibration of the CE being performed between each set of samples. For the 97 samples examined, the pooled standard deviation was 0.3 bp. The observed genotype frequencies determined from the sample set did not deviate significantly from Hardy-Weinberg expectations. From these CE results, we conclude that HUMTH01 PCR products can be accurately and precisely sized by capillary electrophoresis using the method described.
毛细管电泳（CE）是一种分析技术，可对聚合酶链反应（PCR）产生的扩增DNA片段进行快速、高分辨率分析。在本研究中，使用两种内标作为大小标记，以精确界定PCR产物的大小。该技术应用于短串联重复序列位点HUMTH01的PCR产物分型。HUMTH01由五到七个主要等位基因组成，大小范围在179 - 203 bp之间，每个等位基因相隔4 bp。使用这种遗传标记，对一个包含97个个体的群体进行了聚丙烯酰胺凝胶电泳和毛细管电泳检测。两种技术获得了相同的基因型，证明了毛细管电泳在DNA分型应用中的可靠性。DNA分析以每组10个样本的方式进行，每组样本之间对毛细管电泳进行校准。对于所检测的97个样本，合并标准偏差为0.3 bp。从样本集中确定的观察到的基因型频率与哈迪 - 温伯格预期没有显著偏差。根据这些毛细管电泳结果，我们得出结论，使用所述方法通过毛细管电泳可以准确且精确地确定HUMTH01 PCR产物的大小。
