Rapid differentiation of Hanwoo and Holstein meat using multiplex allele specific polymerase chain reaction protocols

Koh, Ba-Ra-Da

Korean Journal of Veterinary Research 2005;45(3):351-357.

Ba-Ra-Da Koh

Gwangju Metropolitan Health & Environment Research Institute

Multiplex allele specific PCR 방법을 이용한 한우고기와 젖소고기의 신속한 판별

고바라다

광주광역시보건환경연구원

Abstract

Here I describe a multiplex allele specific PCR-based approach for the rapid detection between Hanwoo and Holstein meat associated with Melanocortin 1 receptor (MC1R) gene. Specific and universal oligonucleotide primers were used in combination to detect the presence of a single nucleotide polymorphism within the bovine MC1R DNA sequence. The presence of the bovine MC1R gene is indicated by the production of a single control PCR product, whilst positive samples generate an alternative smaller specific product over the same region. The mutations in MC1R104 codon revealed depending on the presence or absence of an indicative fragment amplified from the wild-type allele of this codon. As little as 0.39 ng and 1.56 ng of genomic DNA of Hanwoo and Holstein could be detected by MAS-PCR assay, respectively. This technique, which is widely used in human genetic screening, provides a reliable and sensitive result that has not been documented for the identification of bovine coat color. The MAS-PCR assay approach was proven to be useful in complementing routine beef DNA analysis for differentiation of these MC1R variants and it would facilitate the screening of deceiving sales of Holstein meat in the butcher shop.

Key Words: MC1R gene, coat color, Multiplex allele specific PCR, SNP, Hanwoo