Establish a practical protocol for genetic testing in Phu Quoc ridgeback dog by microsatellite markers

Abstract

Phu Quoc ridgeback dogs are special dog breed of Vietnam, living in Phu Quoc island. Their genome has not been fully known, and now they are facing inbreeding as a result of mating between closely-related individuals. Microsatellite markers are the target sequences for its wide range of applications in genotyping and fingerprinting. In this study, six microsatellite markers (AHT137, FH2054, INRA21, INU030, REN162C04, REN247M23) were chosen for our genetic testing protocol. As we have effectively employed non-denaturing polyacrylamide gel electrophoresis (Non-denaturing PAGE) for genotyping with STR markers, the separation of STRs on non-denaturing PAGE was found easier and economical. By employing non-denaturing PAGE, we were successful to clearly separate SSR alleles with a 2-bp difference in size. In addition, silver staining was successfully employed for staining DNA for its high sensitivity to stain DNA in polyacrylamide gel. Thus, our protocol gives an economical and effective tool for researchers involved in DNA fingerprinting work with microsatellite markers. Keywords: Phu Quoc ridgeback dogs; Microsatellite markers; Polyacrylamide gel electrophoresis (PAGE); denaturing PAGE; non-denaturing PAGE; Silver staining.