DNA molecular identification of Manis pentadactyla.
10.19540/j.cnki.cjcmm.2017.0092
- Author:
Yan YIN
1
;
Xun LIU
2
;
Bing WANG
3
;
Lin-Hui GAO
1
;
Xia-Nan ZHANG
1
Author Information
1. School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
2. Suzhou Vocational Health College, Suzhou 215009, China.
3. Shanxi University of Traditional Chinese Medicine, Taiyuan 030024, China.
- Publication Type:Journal Article
- Keywords:
COⅠ;
Manis pentadactyla;
molecular identification;
Cytb
- From:
China Journal of Chinese Materia Medica
2017;42(11):2078-2084
- CountryChina
- Language:Chinese
-
Abstract:
The study was aimed to establish a stable, accurate site specific PCR identification system to identify Manis pentadactyla and its adulterants using DNA molecular identification. The genomic DNA was extracted from experimental samples using the DNA extraction kit. The Cytb and CO Ⅰ genes were amplified using PCR and sequenced bi-directionally. Obtained sequences were assembled using the BioEdit software. The neighbor-joining tree was constructed by MEGA 6.0. Specific identification primers were designed according to the specific allelets, and PCR reaction system was optimized. The results indicated that the Cytb and CO Ⅰ sequence both were able to be used to identify M. pentadactyla and its adulterants. With the specific primers CO Ⅰ-S10/A5, the M. pentadactyla could be amplified a 400 bp DNA band when the annealing temperature ranged from 55 to 60 ℃ and the amount of DNA template ranged from 3 to 100 ng within 35 PCR cycles. However, other adulterants displayed no relevant bands. So that primers CO Ⅰ- S10 / A5 can be used to identify the M. pentadactyla with the adulterants.
