Specific primer and detection method of macaca thibetana mitochondria D-loop region and application

Abstract

The invention discloses a specific primer and a detection method of macaca thibetana mitochondria D-loop region and application. Primer sequences are shown as SEQ ID No: 1 and SEQ ID No: 2. When the primer is used for PCR amplification, an amplification system is as follows: 2[mu]L of genomic DNA, 5[mu]L of 10*PCR buffer solution, 2[mu]L of Mg<2+> with the concentration of 1.5mM, 2.5[mu]L of Taq enzyme with the concentration of 1U / MuL, 5[mu]L of dNTPs with the concentration of 0.2mM, 1[mu]L of positive and negative primers with the concentration of 10mM respectively and the balance of ddH2O, altogether 50[mu]L; amplification procedures are as follows: predenaturing for 5min at 94 DEG C; denaturing for 30s at 94 DEG C, annealing for 30s at 60 DEG C, stretching for 90s at 72 DEG C, altogether 30 cycles, and finally stretching for 8min at 72 DEG C. The invention designs the specific primer for amplifying a full-length sequence of the macaca thibetana mitochondria D-loop region; a completeD-loop sequence can be rapidly and accurately amplified.

Description

technical field [0001] The invention belongs to the technical field of genetic analysis of Tibetan chief monkeys, and in particular relates to a specific primer, a detection method and an application of the mitochondrial D-loop region of Tibetan chief monkeys. Background technique [0002] Tibetan monkey (Macaca thibetana), also known as Daqing monkey and Tibetan monkey, belongs to the genus Macaca of the family Cercopithecidae (Primates) and is a unique non-human primate species in my country. Sichuan, Guizhou, Anhui, Fujian and Zhejiang in the northern subtropical zone. The Tibetan monkey belongs to the national second-class protected wild animals. It is listed as an appendix 2 species by the Convention on International Trade in Endangered Wild Fauna and Flora (CITES), and is listed as a "near-threatened" species in the Red List of Threatened Species by the International Union for Conservation of Nature (IUCN). Due to human disturbance in recent years, the number of wild T...

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Problems solved by technology

[0004] In view of the above-mentioned deficiencies in the prior art, the present invention provides a specific primer and detection method and application of the mitochondrial D-loop region of the Tibetan chief monkey, which can effectively solve the problem that the primers and detection methods designed in the prior art cannot obtain complete D-loop sequence, in order to obtain a complete D-loop sequence, it is necessary to connect the cloning vector and then perform sequencing. The process is cumbersome and the genetic diversity of the Tibetan monkey population cannot be fully evaluated by using part of the D-loop sequence. question

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