ABSTRACT
For rapid identification of Cervus nippon, C. elaphus and their hybridize samples, the specific PCR for mutual authentication of them was established based on the SNPs in COI and SRY sequence. C. nippon, C. elaphus and their hybridize samples were collected from different origins, total DNA of 24 identified samples were extracted, and the COI and SRY gene was seqenced. SNPs in the COI and SRY sequences of the samples were found by Clustul X 2.1 program. Primers for identifying C. nippon and C. elaphus were designed according to the SNP site, two multi-PCR reaction system were established to identify them. In addition, 24 samples which were randomly collected in different herbal medicine market were identified. The band special for C. nippon (232 bp)and band special for C. elaphus (518 bp) based on COI sequence,and the band special for C. nippon (803 bp)and band special for C. elaphus (425 bp) based on SRY sequence, were found using multi-PCR reaction, and three of the twenty-four samples were identified as the hybridize samples. The multi-PCR reaction system could be used to identify C. nippon, C. elaphus and their hybridize samples.
ABSTRACT
Objective: For rapid identification of Houttuynia cordata and Gymnotheca chinensis, the specific PCR for mutual authentication of them was established based on the SNPs in matK sequence. Methods: H. cordata and G. chinensis samples from different origins were collected, total DNA of all samples was extracted, and the matK gene was seqenced. SNPs in the matK sequences of all the samples were found by ClustulX 2.1 program. Primers for identifying H. cordata and G. chinens were designed according to the SNP site, and specific PCR method was established to identify them, for rapid detection by addition of SYBR Green I dye. In addition, constructing a multi-PCR reaction system, and then the PCR reaction system was optimized. Results: The band special for H. cordata (185 bp) and band special for G. chinensis (389 bp) were found using specific PCR reaction and multi-PCR reaction, and SYBR Green I dye can be used for rapid detection. Conclusion: The multi-PCR reaction system could be used to identify H. cordata and G. chinensis.