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Primer and method for evaluating biomass of trachemys scripta elegans based on environmental DNA (eDNA) technology

A red-eared turtle biomass technology, applied in the field of primers for red-eared turtle biomass assessment, can solve the problems of low efficiency, unfavorable red-eared turtle invasion in the wild, time-consuming and labor-intensive problems

Active Publication Date: 2018-09-04
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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AI Technical Summary

Problems solved by technology

Traditional turtle distribution and population surveys are mainly based on field animal entity surveys, which are time-consuming, laborious, and inefficient, and are not conducive to quickly grasping the wild invasion status of red-eared turtles on a large scale

Method used

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  • Primer and method for evaluating biomass of trachemys scripta elegans based on environmental DNA (eDNA) technology
  • Primer and method for evaluating biomass of trachemys scripta elegans based on environmental DNA (eDNA) technology
  • Primer and method for evaluating biomass of trachemys scripta elegans based on environmental DNA (eDNA) technology

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Experimental program
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Effect test

Embodiment 1

[0026] (1) Primer design

[0027] qPCR primers were designed according to the COI sequence of the mitochondrial DNA gene of the red-eared turtle, specifically:

[0028] The upstream primer F is: 5'-GGGATGACCAAATCTACAATG-3';

[0029] The downstream primer R is: 5'-TAATGGCACGAGTCAGTTCC-3'.

[0030] (2) Standard curve making of red-eared turtle DNA concentration and Ct value

[0031] The leg muscle tissue of the red-eared turtle was taken, and the DNA of the red-eared turtle was extracted according to the instructions of the animal tissue genomic DNA extraction kit. The DNA concentration was measured by a NanoDrop 3000 fluorescence spectrophotometer, and the concentration of the DNA stock solution was 0.248 μg / μl. The red-eared turtle DNA stock solution was diluted 8 times, 16 times, 32 times, 64 times, 128 times, 256 times, 512 times, 1024 times, 2048 times in sequence, including 10 DNA concentrations in the stock solution. The sequentially diluted DNA was used as the reactio...

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Abstract

The invention discloses a primer and a method for evaluating the biomass of trachemys scripta elegans based on an environmental DNA (eDNA) technology. The method comprises the following steps: 1) preparing a standard curve of DNA concentration and Ct value of the trachemys scripta elegans by a qPCR technology; 2) measuring the eDNA concentration of the trachemys scripta elegans in a water body under different trachemys scripta elegans stocking density environments according to the standard curve; 3) making a corresponding curve of the measured eDNA concentration and the corresponding biomass of the trachemys scripta elegans. The results show that the eDNA concentration and the biomass of the trachemys scripta elegans is of a linear relation, and R2 is 0.9429. The method provided by the invention can quickly identify the trachemys scripta elegans by detecting a water body in laboratory environment and evaluate the biomass of the trachemys scripta elegans according to the eDNA concentration, so as to provide technical support to the field distribution investigation, quantity evaluation and invasion degree and grade evaluation and the like of the trachemys scripta elegans.

Description

Technical field: [0001] The invention relates to the field of biotechnology, in particular to a primer and a method for evaluating the biomass of red-eared turtles based on environmental DNA technology. Background technique: [0002] Environmental DNA (environmental DNA, eDNA) refers to a collection of all DNA fragments directly extracted from environmental samples (such as water, feces, soil, excrement, etc.). The method of qualitative or quantitative analysis of organisms in the environment by directly extracting DNA fragments from environmental samples and using various molecular biological methods is called eDNA technology. In the research of aquatic animals, compared with traditional survey methods, eDNA technology has the characteristics of high efficiency, rapidity, high sensitivity and no damage to organisms. In recent years, eDNA technology has been widely used in aquatic biological invasion, rare species monitoring, biodiversity assessment and biomass estimation. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q2563/107C12Q2531/113
Inventor 龚世平曹代男魏玉峰李伟业段好冉葛研
Owner INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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