The analysis of simple sequence repeat DNA in soybean by Capillary Gel Electrophoresis.

Abstract

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.