Development of a 17-plex STR typing system for the identification of individuals and parentage testing in cattle

Abstract

Accurate identification of animals and the verification of their parentage can be used to pedigree populations and support selective breeding. The International Society for Animal Genetics recommended 16 cattle STRs for individual identification and parentage testing in cattle, but no multiplex STR typing system contains these 16 STRs. Here, we develop an efficient 17-plex multiplex typing system for cattle that contains the 16 ISAG recommend STRs and a sex-determining marker. Compared to the Bovine Parenting Typing Kit (containing 11 of the 16 ISAG recommend STRs), our new typing system not only increases the number of molecular markers, but also simplifies the PCR operation and shortens the time for the typing procedure (from 4.5 h to 1 h 37 min). Profile can be generated from a single PCR reaction using as little as 1 ng of DNA. The combined probabilities of paternity exclusion CPE_duo and CPE_trio were 0.999804697 and 0.999999260, respectively. These results indicate that our 17-plex typing system is a fast, sensitive and species-specific method for the identification of individuals and their parentage for cattle. The application of this system will improve the efficiency of the identification of cattle individuals and their paternity, supporting population genetic research and the selective breeding of cattle.

Keywords: Cattle; Microsatellite; Multiplex PCR; Parentage testing.

Fig. 1

Genotype profile of a cattle generated by the 17-plex typing system.

Fig. 2

Sensitivity testing for the 17-plex PCR system using a range of DNA templates. Black arrows indicate allele peaks that were below the 200 rfu threshold and red arrows indicate nonspecific peaks using low DNA templates.