A rapid and simple method for sex identification by heteroduplex analysis,using denaturing high-performance liquid chromatography (DHPLC)

A novel method for sex identification. using a denaturing high-performance liquid chromatography (DHPLC) system, is described. Among many methods for identifying sex, the most popular and credible system has been the polymera se chain reaction (PCR) method, using nucleotide primer sets of the ameloge nin gene, which is shared on both the X and Y chromosomes. With this conven tional method, the judgment depends on detection of the size difference bet ween the PCR products derived from the X and Y chromosomes. In this study, we adopted DHPLC to detect the difference by checking heteroduplex formatio n between the products, which enabled us to shorten the PCR products to 45 bp and the separation time to within a period of 8min per sample. This new system may have wide applications in many different fields, such as forensi c medicine, prenatal diagnosis, inbreeding of animals, and anthropology.