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Homogeneous fluorescent specific PCR for the authentication of medicinal snakes using cationic conjugated polymers
Chao Jiang1,2, Yuan Yuan1, Libing Liu3
1State Key Laboratory Breeding Base of Dao-di Herbs, National Resources Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, P.R. China.
Abstract:
A label-free, homogenous and sensitive one-step method for the molecular authentication of medicinal snakes has been developed by combining a rapid PCR technique with water-soluble cationic conjugated polyelectrolytes (CCPs). Three medicinal snake materials (Deinagkistrodon acutus, Zaocys dhumnades and Bungarus multicinctus; a total of 35 specimens) and 48 snake specimens with similar morphologies and textures were clearly distinguished by the naked eye by utilizing a CCP-based assay in a high-throughput manner. The identification of medicinal snakes in patented Chinese drugs was successfully performed using this detection system. In contrast to previous fluorescence-labeled oligonucleotide detection and direct DNA stain hybridization assays, this method does not require designing dye-labeled primers, and unfavorable dimer fluorescence is avoided in this homogenous method.
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