Rare species DNA information acquiring method

An acquisition method and species technology, applied in the field of DNA information acquisition of rare species, can solve the problems of difficulty in obtaining financial support and high cost, and achieve the effect of expansion and application

Inactive Publication Date: 2018-10-30
ZHEJIANG FORESTRY UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

With the development of omics research technology, it is not difficult to obtain the DNA information of any organism, but the cost is too high, and it is difficult to obtain any level of financial support without a certain basis for preliminary work

Method used

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  • Rare species DNA information acquiring method

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Embodiment Construction

[0019] In this embodiment, taking the South China tiger as an example, the following methods were tried and succeeded.

[0020] (1) After the South China tiger tissues were obtained, the DNA was purified by phenol / chloroform extraction, and the DNA samples were subjected to intermittent vortex shaking for 20 s to randomly fragment the DNA.

[0021] (2) Through 0.8% agarose gel electrophoresis (add 0.8 g agarose to 100 mL 1×TAE buffer solution, heat it in a microwave oven to melt it completely, and add a small amount of ethidium bromide when the temperature drops to around 50°C Mix well and pour into a gel-making mold) to separate the DNA and recover the DNA with a molecular weight between 500-1 000bp. The specific method is as follows:

[0022] Prepare the centrifuge tube for weighing, cut the gel containing the DNA sample under the ultraviolet light, put it into the centrifuge tube and weigh it, calculate the mass of the gel, then purify the DNA according to the instructions...

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Abstract

The invention relates to a rare species DNA information acquiring method, and belongs to the technical field of measuring or detecting methods containing enzyme or microorganisms. The method comprisesthe following steps: after acquiring a tissue of a south China tiger, purifying DNA, and enabling the DNA to generate random fragmentation, separating the DNA, and recovering, performing a A-adding catalytic reaction on the recovered DNA, and performing a coupled reaction on a T-carrier, performing conversion on a coupled product, screening a positive bacterial colony, culturing overnight, performing plasmid recovery, sequencing, and performing a specific primer design, synthetizing, and implementing PCR again and sequencing a PCR product, performing comparison with a sequence used in the primer design, if a result is consistent, it is demonstrated that the DNA fragment is the DNA of the target species surely. The method is used for acquiring DNA information of the rare species, and has the advantages of simple and convenient operation, high acquiring rate, low cost and the like.

Description

technical field [0001] The application relates to a method for acquiring DNA information of rare species, which belongs to the technical field of measurement or detection methods involving enzymes or microorganisms. Background technique [0002] South China Tiger ( Panthera tigris Amoyensis ) is a subspecies of tiger unique to my country. It was included in the CITES Appendix I protection list in 1981, also known as "Chinese tiger". It is currently considered extinct in nature and is only kept in a few limited zoos for tourists to watch . There are many such endangered animals in the world. With the development of omics research technology, it is not difficult to obtain DNA information of any organism, but the cost is too high, and it is difficult to obtain any level of financial support without a certain basis for preliminary work. Therefore, how to obtain some DNA information and carry out some work in the case of limited funds is particularly important to do a good job ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C12N15/10C12N15/70G06F19/24
CPCC12N15/1003C12N15/70C12Q1/6869G16B40/00
Inventor 杨仙玉张先福马敬华刘建勋施伟斌
Owner ZHEJIANG FORESTRY UNIVERSITY
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