Establishment and Application of a Fluorescent Polymerase Chain Reaction−Restriction Fragment Length Polymorphism (PCR-RFLP) Method for Identifying Porcine, Caprine, and Bovine Meats

A method of fluorescent Polymerase Chain Reaction−restriction fragment length polymorphism (PCR-RFLP) was applied as an analytical and quantitative tool for meat identification. Following alignments of the nucleotide sequences, an oligonucleotide primer pair was designed to amplify the partial seque...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2003-03, Vol.51 (7), p.1771-1776
Main Authors: Sun, Yu-Ling, Lin, Chich-Sheng
Format: Article
Language:English
Subjects:
Animals
autoclaving
Base Sequence
beef
Biological and medical sciences
capillary electrophoresis
cats
cattle
Cattle - genetics
chickens
cooking
DNA fingerprinting
dogs
ducks
fish
fluorescence
Fluorescent Dyes
Food industries
Fundamental and applied biological sciences. Psychology
geese
genes
goats
Goats - genetics
Meat - classification
Meat and meat product industries
mitochondrial DNA
Molecular Sequence Data
nucleotide sequences
polymerase chain reaction
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
pork
ribosomal RNA
RNA - analysis
RNA - chemistry
RNA, Ribosomal - analysis
RNA, Ribosomal - chemistry
species identification
swine
Swine - genetics
turkeys
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Summary:A method of fluorescent Polymerase Chain Reaction−restriction fragment length polymorphism (PCR-RFLP) was applied as an analytical and quantitative tool for meat identification. Following alignments of the nucleotide sequences, an oligonucleotide primer pair was designed to amplify the partial sequences within the 12S ribosomal RNA (12S rRNA) gene of mitochondrial DNA from porcine, caprine, and bovine meats. No fragment can be amplified from dog, cat, fish, duck, goose, turkey, and chicken DNA with the primer pair. Using fluorescence sensor capillary electrophoresis, the species-specific DNA fingerprints of pork, goat, and beef were generated by restriction enzyme digestion following a fluorescence-labeling PCR amplification. Species identification was conducted on the meat mixtures. The reliably semiquantitative levels were below 1% for binary mixtures of pork, goat, and beef. Cooking and autoclaving of meats did not influence the generation of the PCR-RFLP profiles or the analytical accuracy. Keywords: Meat identification; PCR-RFLP; mitochondrial DNA; 12S ribosomal RNA gene
ISSN:0021-8561
1520-5118
DOI:10.1021/jf020860u