Conventional nucleic acid amplification methods are reliable, but the requirement of complex equipment, skilled technicians and long operation time limit their on-site use. Here, a simple denaturation bubble-mediated strand exchange amplification method (SEA) requiring only a pair of primers and one polymerase was first reported for identifying adulteration of pork source by targeting the species-specific mitochondrial DNA sequence. The SEA method could detect 1% pork total DNA by both colorimetric and fluorescence determination. The whole detection process could be finished within 1 hour by coupling with fast tissue DNA extraction method, only requiring a simple heating block, and hence making it suitable for on-site meat species identification.
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