Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Molecular marker for identifying sika deer Sichuan subspecies, identification method and application

A molecular marker, sika deer technology, applied in the field of molecular biology, achieves high efficiency, high accuracy, and good stability

Pending Publication Date: 2022-08-02
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are no related reports on the identification of Sichuan subspecies of sika deer using molecular markers

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Molecular marker for identifying sika deer Sichuan subspecies, identification method and application
  • Molecular marker for identifying sika deer Sichuan subspecies, identification method and application
  • Molecular marker for identifying sika deer Sichuan subspecies, identification method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, for identifying the acquisition of the specific SNP marker of sika deer Sichuan subspecies

[0024] 1. Screening of sika deer Sichuan subspecies-specific SNP loci

[0025] Combined with the measured mtDNA sequences of different subspecies samples of sika deer, such as Northeast Asia, Sichuan, South China, Taiwan, Hokkaido, Honshu, Nominative and Yakushima, the Mega6.0 was used to compare the samples. For the analysis, we mainly screened loci common to the same subspecies but specific to other subspecies, and screened 5 differential SNP loci, such as figure 1 and shown in Table 1. The five marker sites are all located on the 306bp fragment of the ATP6 gene of the sika deer mtDNA genome; the marker S-1 is at the 31 bp, which is the base T in the Sichuan subspecies, and the base C in the non-Sichuan subspecies; the marker S-2 is located at the 73 bp, Sichuan The base C in the subspecies is not the base T in the Sichuan subspecies; the marker S-3 is at the ...

Embodiment 2

[0032] Embodiment 2, the establishment of the method for identifying sika deer Sichuan subspecies

[0033] Using the primer pairs shown in the above-mentioned SEQ ID NO.1-SEQ ID NO.2, a PCR experiment was carried out to establish a method for identifying the Sichuan subspecies of sika deer. The PCR amplification conditions are shown in Table 2.

[0034] Table 2 PCR amplification conditions

[0035]

[0036] The PCR reaction system is shown in Table 3.

[0037] Table 3 PCR system

[0038]

[0039] The PCR amplification process was as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 sec, annealing at 58°C for 30 sec, extension at 72°C for 30 sec, a total of 30 cycles; extension at 72°C for 5 min, and storage at 4°C.

[0040] Take 3.5 μL of PCR product and detect it in 1% agarose gel electrophoresis. The electrophoresis result ( figure 2 ) is consistent with the target fragment, and the band is single and dense, which can be used for subsequent seq...

Embodiment 3

[0043] Embodiment 3. the concrete application of the method that the present invention identifies sika deer Sichuan subspecies

[0044] A total of 120 samples collected from China and Japan were selected, and genomic DNA was extracted respectively. Experimenter A randomly mixed these samples and renumbered them. Experimenter B used the primers designed in the present invention to carry out PCR amplification of these samples, and the amplified products Sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing. View the sequencing peak map through Bioedit 7.0. The peaks and troughs of the sequencing results are separated, without overlap, and without misreading, indicating that the sequencing results are credible. The sequences were aligned with Mega6.0, and the bases of the specific SNP sites of sika deer Sichuan subspecies were used as the judgment basis.

[0045] It is determined according to the mtDNA-specific SNP molecular marker determined in the present invention...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a molecular marker for identifying sika deer Sichuan subspecies, an identification method and application. In order to solve the problem that no molecular marker and identification method for identifying sika deer Sichuan subspecies exist in the prior art, the sika deer molecular marker is deeply studied, the SNP molecular marker for identifying the sika deer Sichuan subspecies is obtained, and the molecular marker comprises five SNP marker sites. The five SNP markers are positioned on an ATP6 gene segment, and have the characteristics of strong specificity and good stability. The invention also provides a pair of SNP marker primers for identifying sika deer Sichuan subspecies, and establishes an accurate and reliable detection method. The molecular marker overcomes the defect that no molecular marker detection technology for identifying the sika deer subsp.Sichuan does not exist in the field at present, accurate identification of the sika deer subsp.Sichuan is achieved, and the molecular marker has important theoretical and application value in the aspects of protection and management of sika deer resources.

Description

technical field [0001] The invention relates to a molecular marker, an identification method and an application for identifying sika deer Sichuan subspecies, and belongs to the technical field of molecular biology. Background technique [0002] The sika deer (Cervus nippon) belongs to Mammalia, Artiodactyla, Cervidae and Cervus. At present, it is generally accepted that the sika deer are divided into 13 subspecies. The 6 existing subspecies in Japan are the Hokkaido subspecies (Cervusnipponyesoensis), the Honshu subspecies (Cervus nippon centralis), the named subspecies (Cervusnipponnippon), and the Yakushima subspecies. (Cervus nippon yakushimae), horsehair subspecies (Cervusnipponmageshimae) and Ryukyu subspecies (Cervus nipponkeramae). Vietnam has 1 subspecies: the Vietnamese subspecies (Cervus nippon pseudaxis). There are 6 subspecies of Chinese sika deer, of which the North China subspecies (Cervusnipponmandarinns) and the Shanxi species (Cervus nippon grassianus) are...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 邢秀梅董依萌王天骄刘华淼李洋张然然
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com