Molecular identification method of saurogobio dabryi and saurogobio dumerili based on COI gene segment

A technique for long snake globber and snake globber, which is applied in the field of genetics to achieve the effect of good stability and high repetition rate

Pending Publication Date: 2020-10-02
FRESHWATER FISHERIES RES INSITUTE OF JIANGSUPROVINCE
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the use of Cytochrome Oxidase subunit I (COI) gene as a detection target for the identification of scorpion and long snake.

Method used

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  • Molecular identification method of saurogobio dabryi and saurogobio dumerili based on COI gene segment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Primer design

[0026] Design a pair of specific PCR amplification primers, the primer sequence is: F: 5'-TCAACCAACC-ACAAAGACATTGGCAC-3' (SEQ ID NO.1), and the downstream primer is R: 5'-TAGACTTCTG-GGTGGCCAAAGAATCA-3' (SEQ ID NO.2).

[0027] 2. Sample collection

[0028] Take 40 individuals of Snake Snake and Snake Snake to be identified.

[0029] 3. Genomic DNA extraction

[0030] Genomic DNA of Snake and Long Snake was extracted by genome extraction kit, and the DNA was dissolved in TE solution. DNA integrity was identified by 1.0% agarose gel electrophoresis, the OD value was measured by ultraviolet spectrophotometer and diluted to 100ng / μL, and stored at -20°C for future use.

[0031] 4. PCR amplification and detection

[0032] The extracted DNA was used as a template, and the above primers (F and R) were used for PCR amplification. The PCR reaction system is 50 μL: 1 μL template DNA (100 ng / μL); 25 μL PCR mixture (1.25 IU Taq enzyme, 20 mmol / L Tris-HCl, pH...

Embodiment 2

[0037] According to the method in Example 1, the sample was expanded to 80 individuals of the scorpion and the long-snake, and the results still showed that the base of the fourth site (COI-4) of the COI gene of the Snake was T, and the fourth site of the long-snake (COI-4) base is C.

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Abstract

The invention discloses a molecular identification method of saurogobio dabryi and saurogobio dumerili based on a COI gene sequence. The method comprises the following methods of (1) respectively extracting genome DNA of the saurogobio dabryi and the saurogobio dumerili; (2) performing PCR amplification on cytochrome oxidase subunit I genes of the saurogobio dabryi and the saurogobio dumerili; and(3) after amplification products are purified, performing direct sequencing, wherein a COI gene amplification product of the saurogobio dabryi is shown as SEQID NO.3, and a COI gene amplification product of the saurogobio dumerili is shown as SEQID NO.4. The saurogobio dabryi lies in that fourth-site base of the COI gene sequence is T, and the saurogobio dumerili lies in that fourth-site base ofthe COI gene sequence is C. According to the method, the saurogobio dabryi and the saurogobio dumerili can be simply, quickly and effectively identified, the result stability is good, the repetition rate is high, the blank that at current, identifying the saurogobio dabryi and the saurogobio dumerili by a domestic molecular biology standard is filled, and development of the resource investigationand fishery resource species diversity and heredity diversity protection on the saurogobio dabryi and the saurogobio dumerili can be facilitated.

Description

technical field [0001] The invention belongs to the field of genetics, and relates to a method for molecular identification of scorpion and longsnake based on COI gene fragments. Background technique [0002] Saurogobio dabryi and Saurogobio dumerili are two small fishes commonly found in freshwater in my country. They belong to Cyprinidae and belong to the genus Saurogobio dabryi. They often inhabit the middle and lower waters of rivers and lakes. Snake scorpion and long snake scorpion are omnivorous fish, widely distributed and large in number, and they are common species in fish community structure. Both fish species are edible and have certain economic value. Over the years, due to unfavorable factors such as overfishing, environmental pollution, and construction of water conservancy projects, the habitat of fish has been destroyed, and the amount of fishery resources has declined sharply. It is imminent to strengthen the protection of fishery resources. The investigat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 李大命钟立强林海赵沐子朱凛朱莉张可李春宁
Owner FRESHWATER FISHERIES RES INSITUTE OF JIANGSUPROVINCE
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