DNA mini-barcoding: An innovative solution for degraded DNA and its multidisciplinary application landscape

DNA mini-barcoding amplifies short DNA fragments, overcoming the inherent limitations of conventional barcoding that targets longer regions and frequently fails in highly degraded DNA. Capitalizing on its distinct advantages in analyzing highly fragmented and complex-matrix DNA, this technology has emerged as a robust approach across disciplines. This review synthesizes current advances and delineates its expanding application landscape. Specifically, it facilitates biological evidence tracing in forensic science, enables the identification of fossils and archaeological remains, and verifies food authenticity. Furthermore, the technology permits the precise detection of adulteration within the traditional Chinese medicine trade chain and allows for effective species identification from environmental samples for biomonitoring. Importantly, it also shows promise in biodiversity assessment and the regulation of wildlife trade, especially when integrated with complementary techniques for trace DNA detection. This review elucidates the technical principles, evaluates interdisciplinary progress, and provides methodological references. Future research should prioritize optimizing primers for extreme degradation, constructing comprehensive reference libraries, and integrating mini-barcoding with quantitative technologies to enhance diagnostic precision.