He Zhi-Yi, Tang Xian-Ming, Liu Jian-Hui, Yuan Yuan, Jiang Chao, Zhao Yu-Yang, Wang Yang
Harbin Food and Drug Inspection Center, Harbin 150525, China.
State Key Laboratory Breeding Base of Dao-di Herbs，National Resources Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijng 100700, China.
Zhongguo Zhong Yao Za Zhi. 2017 Jul;42(13):2467-2472. doi: 10.19540/j.cnki.cjcmm.20170614.012.
Rapid allele-specific PCR primer was designed base on Cytb 155 A/T single nucleotide polymorphism, DNA was extracted by alkaline lysis and the PCR reaction systems including denatured and annealing temperature and cycle numbers were optimized. The results were performed to authenticate Ranae Oviductus and its 4 adulterants. When 100×SYBR Green I was added in the PCR product at 90 ℃ denatured 3 s, 62 ℃ annealing 20 s and 32 cycle. Ranae Oviductus visualized strong green fluorescence under 365 nm UV lamp whereas adulterants appeared negative. The whole process can be completed in 40 minutes．The established method provides the technical support for authentication of the Ranae Oviductus.
