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Primer for amplifying marten species cytochrome b gene and method for identifying sable and pine marten

A cytochrome, b gene technology, applied in biochemical equipment and methods, determination/inspection of microorganisms, DNA/RNA fragments, etc., can solve problems such as easy misjudgment

Inactive Publication Date: 2010-08-04
NORTHEAST FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to solve the problem that existing sable and pine marten morphological identification methods are easily misjudged, and provide a primer for amplifying the cytochrome b gene of the sable species and a method for discriminating sable and pine marten

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  • Primer for amplifying marten species cytochrome b gene and method for identifying sable and pine marten
  • Primer for amplifying marten species cytochrome b gene and method for identifying sable and pine marten
  • Primer for amplifying marten species cytochrome b gene and method for identifying sable and pine marten

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specific Embodiment approach 1

[0010] Specific embodiment 1: In this embodiment, the primers for amplifying the cytochrome b gene of the mink species are composed of an upstream primer and a downstream primer, wherein the upstream primer is Mz-L: 5'-CATCGGAACCAGTCTTGTAG-3', and the downstream primer is Mz-H: 5'-GTTGGCGGGGATGTAGTTGTC-3'.

specific Embodiment approach 2

[0011] Specific embodiment two: the method for identifying sable and pine marten in this embodiment is carried out according to the following steps: one, get the biological sample material of the sample to be tested, adopt the DNA extraction kit to extract the biological sample material DNA, and then use the amplified mink species The primers of cytochrome b gene were used for PCR amplification, and 20 μL of PCR reaction system was prepared with a final concentration of 0.2 μmol L -1 The upstream primer, the final concentration is 0.2 μmol L -1 The downstream primers, the final concentration is 0.16mmol L -1 dNTP, the final concentration is 1.6mmol·L -1 MgCl 2 , 1.0U of EX-Taq DNA polymerase, 100ng of biological sample DNA and the rest of sterilized deionized water. The PCR reaction program is: 94°C preheating for 3min, 94°C denaturation for 30s, 59°C renaturation for 45s, 72°C Extend 1 min at ℃, cycle 30-35 times and extend at 72 ℃ for 5 min. The target fragment is 316bp i...

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Abstract

The invention discloses a primer for amplifying marten species cytochrome b gene and a method for identifying sable and pine marten, and relates to a primer and a species identifying method. The method solves the problem that the conventional morphologic identification method for the sable and the pine marten easily causes error judgment. The primer consists of an upstream primer and a downstream primer. The method comprises the following steps: 1, extracting DNA, and then performing PCR amplification; 2, performing enzyme digestion; and 3, performing electrophoresis detection. The skin, skeleton, muscle and claw of an animal are used as biological detection materials and then detected by using the identifying method of the invention, and the detection result is completely accurate. The method of the invention not only can be used for wild animal law enforcement, but also can be applied in commercial inspection.

Description

Technical field [0001] The present invention involves a method of primer and species. Background technique [0002] Shobi is a precious fur. In the past, it was one of the types of traditional export fur. At present, due to the scarce population, the Chinese National Wildlife Protection Law has listed sable as a national I -level key protection animal.The pine marten is a proximity species of sable. It is distributed in Europe. It is very similar in shape to the sable, making it difficult to distinguish the shape of sable and pine mink, which has caused some difficulties for related wildlife law enforcement and commodity inspection.The traditional form science identification method is adopted, because the human error is large, it is easy to misjudge, and the accuracy rate is low. Invention content [0003] The purpose of the present invention is to solve the problems that the existing divases and pine marten forms are easy to misjudge, and it provides a method for amplifying the...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 李波徐艳春
Owner NORTHEAST FORESTRY UNIVERSITY
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