Journal
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 75, Issue 9, Pages 1639-1643Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1271/bbb.110024
Keywords
real-time PCR; reference molecule; cytochrome b; meat
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Funding
- Ministry of Health, Labour and Welfare of Japan
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A standard plasmid was constructed as a novel reference molecule for use in real-time quantitative PCR assays to verify the identity of beef, pork, chicken, mutton, and horseflesh. The plasmid contained a target domain of the cytochrome b (cyt b) gene and an artificial DNA sequence. Primers CO-F and CO-R, and probe CO-P were specifically designed to detect the artificial sequence. The calculated R-2 values of the standard curves (10(3)-10(7) copies per reaction) for the five species ranged between 0.998 and 0.999 in the quantification analysis. The constructed plasmid provides a universal method for measuring the copy number of cyt b DNA in minced meat. This method would be a useful procedure for verifying food labels.
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