Species characterization of animal by DNA hybridization

Lee, Myoung-heon; Kim, Sang-keun; Jung, Gab-soo; Park, Jong-myoung

Korean Journal of Veterinary Research 1999;39(3):513-522.

Myoung-heon Lee¹, Sang-keun Kim¹, Gab-soo Jung², Jong-myoung Park²

¹College of Veterinary Medicine, Chungnam National University
²National Veterinary Research & Quarantine Service

DNA hybridization을 이용한 축종특이성 구명

이명헌¹, 김상근¹, 정갑수², 박종명²

¹충남대학교 수의과대학
²국립수의과학검역원

Abstract

DNA hybridization assay using probes prepared from liver was carried out to identify species characterization of the domestic animals. Gel electrophoresis showed that the target DNA extracted from raw muscle were 1kb and uniform pattern while fragments size of heated muscle were irregular. Hybridization was performed by adding 200ng/ml probe in hybridization solution and incubating for 12 hours at $68^{circ}C$. To obtain good discrimination, applied washing buffer and washing step differently depending on the species. The probes of pig, horse and dog formed the specific hybrids with each target DNA respectively. Although cross reaction was detected in cattle, goat and sheep but signal intensity among these species made the discrimination possible each other. Such pattern was the same in the cases of chicken, turkey and duck. The hybridization pattern of heated muscle was similar to that of raw muscle in general, but the signal intensity was inferior to that of raw muscle. Species identification between closely related animal species, hybridized using the target DNA of such closely related animal species as a blocking agent, remarkable increase of discrimination from the evident decrease of non specific reaction compared with the control group. In addition, in the admixture where certain meat was included in the beef, pork, chicken meat, we could find whether any unjust meat was admixed or not. In this case, detection limit of certain meat in admixture was 1%.

Key Words: species characterization, DNA hybridization, muscle