Marino M A, Turni L A, Del Rio S A, Williams P E, Cregan P B
DNA Technologies Development Branch, AFDIL, AFIP, Washington, D.C. 20306-6000, USA.
Appl Theor Electrophor. 1995;5(1):1-5.
The objective of this work is to examine the presence of simple sequence repeat (SSR) DNA in soybean plant genotypes by Capillary Gel Electrophoresis (CGE). The SSR DNA length polymorphism in soybean determines the variation in polymerase chain reaction (PCR) product lengths. Loci were chosen where amplification produced one PCR product per genotype (M.S. Akkaya et al., 1992). The F1 hybrids of parents carrying different alleles produced two PCR products identical to the two parents. The CGE system used a 3%T,3%C polyacrylamide gel capillary with an effective length of 40 cm. The PCR products with lengths of 150 to 200 base pairs were monitored at 260 nm. The analysis time was under 50 minutes. CGE is capable of separating these PCR products by base pair number the same as conventional sequencing gel techniques. CGE offers an automated, high speed, high resolution analytical method for determining soybean SSR allele sizes as compared with the traditional methodologies.
本研究的目的是通过毛细管凝胶电泳（CGE）检测大豆植物基因型中简单序列重复（SSR）DNA的存在情况。大豆中SSR DNA长度多态性决定了聚合酶链反应（PCR）产物长度的变化。选择的基因座上每个基因型的扩增产生一个PCR产物（M.S. Akkaya等人，1992年）。携带不同等位基因的亲本的F1杂种产生了两个与两个亲本相同的PCR产物。CGE系统使用有效长度为40 cm的3%T、3%C聚丙烯酰胺凝胶毛细管。长度为150至200个碱基对的PCR产物在260 nm处进行监测。分析时间在50分钟以内。与传统测序凝胶技术一样，CGE能够按碱基对数分离这些PCR产物。与传统方法相比，CGE为确定大豆SSR等位基因大小提供了一种自动化、高速、高分辨率的分析方法。
