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Method for quickly and efficiently detecting sepiella maindroni population

A high-efficiency technology for squid mansoni, which is applied in the field of rapid and efficient detection of squid mansoni populations, can solve the problems of high cost, damage to target species, time-consuming and labor-intensive, etc., to reduce detection costs, low cost, and improve efficiency and accuracy effects

Inactive Publication Date: 2018-06-26
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the traditional monitoring of the population distribution of the squid mansoni, the on-site survey method is mainly used, such as using nets to collect catches, which is time-consuming, laborious, high in cost, low in catch rate, and may harm the target species or destroy the survey point. However, there are no problems such as rapid and effective detection methods for squid mansoni populations without damaging the environment in the prior art. A rapid and efficient method for detecting the population of squid mansoni by using environmental DNA to detect the population of squid mansoni

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A method for quickly and efficiently detecting the population of squid mansoni, the method for quickly and efficiently detecting the population of squid mansoni comprises the following steps:

[0030] 1) Extract the eDNA sample of the environment to be tested. The collected sample is soil, which is pretreated. The pretreatment step is to place the eDNA sample in liquid nitrogen for sufficient grinding;

[0031] 2) Extract the DNA from the pretreated eDNA sample with a DNA extraction kit, and dissolve the extracted DNA in 100 μL of water to obtain a DNA sample solution;

[0032] 3) For the DNA sample solution, qPCR reaction was carried out with special primers and probes, the special primers were the upstream primer SEQ ID NO.1: 5'-CACCAGACATAGCCTTCC-3' and the downstream primer SEQ ID NO.2:5 '-GCCAGCATGAGATAGATTAC-3'; the probe sequence is SEQ ID NO.3: 5'-HEX-TGTTCATCCAGTTCAGCACCT-TAMRA-3';

[0033] The composition of the qPCR amplification reaction system is: 10 μL of...

Embodiment 2

[0037] A method for quickly and efficiently detecting the population of squid mansoni, the method for quickly and efficiently detecting the population of squid mansoni comprises the following steps:

[0038] 1) Extract the eDNA sample of the environment to be tested. The collected sample is sediment, which is pretreated. The pretreatment step is to place the eDNA sample in liquid nitrogen for full grinding;

[0039] 2) Extract the DNA from the pretreated eDNA sample with a DNA extraction kit, and dissolve the extracted DNA in 100 μL of water to obtain a DNA sample solution;

[0040] 3) For the DNA sample solution, qPCR reaction was carried out with special primers and probes, the special primers were the upstream primer SEQ ID NO.1: 5'-CACCAGACATAGCCTTCC-3' and the downstream primer SEQ ID NO.2:5 '-GCCAGCATGAGATAGATTAC-3'; the probe sequence is SEQ ID NO.3: 5'-HEX-TGTTCATCCAGTTCAGCACCT-TAMRA-3';

[0041] The composition of the qPCR amplification reaction system is: 10 μL of P...

Embodiment 3

[0045] A method for quickly and efficiently detecting the population of squid mansoni, the method for quickly and efficiently detecting the population of squid mansoni comprises the following steps:

[0046] 1) Extract the eDNA sample of the environment to be tested. The collected sample is water, which is pretreated. The pretreatment step is to place the eDNA sample filter membrane in alcohol for full immersion preservation;

[0047] 2) Extract the DNA from the pretreated eDNA sample with a DNA extraction kit, and dissolve the extracted DNA in 100 μL of water to obtain a DNA sample solution;

[0048] 3) For the DNA sample solution, qPCR reaction was carried out with special primers and probes, the special primers were the upstream primer SEQ ID NO.1: 5'-CACCAGACATAGCCTTCC-3' and the downstream primer SEQ ID NO.2:5 '-GCCAGCATGAGATAGATTAC-3'; the probe sequence is SEQ ID NO.3: 5'-HEX-TGTTCATCCAGTTCAGCACCT-TAMRA-3';

[0049] The composition of the qPCR amplification reaction sy...

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PUM

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Abstract

The invention relates to a method for quickly and efficiently detecting a sepiella maindroni population by using environmental DNA to detect the sepiella maindroni population without damaging the environment and the species population. Collected eDNA samples are taken from the soil, sediment and water and the like, no environmental burden is caused to detection points, and no adverse influence iscaused to the living environment of the species. Special primers and probes are utilized to accurately, quickly and efficiently detect the DNA of sepiella maindroni, the efficiency and accuracy of thedetection are improved, and the detection cost is reduced. Compared with a field survey method used by the transmission of the sepiella maindroni population distribution detection, the method has theadvantages of saving time and labor, being low in cost and efficient and causing no harm to target species and no damage to a survey point ecosystem.

Description

technical field [0001] The invention relates to a method for detecting the population of squid mansoni, in particular to a method for detecting the population of squid mansoni quickly and efficiently by adopting environmental DNA without damaging the environment and species population. Population method. Background technique [0002] Sepiella japonica is an annual medium-sized squid. Cephalopoda, Sepiaceae. Commonly known as cuttlefish, it grows fast and has delicious meat. It is a popular seafood food in the market. Mansoni squid is mainly distributed in the Yellow Sea and East China Sea in my country, and is a close-range migratory species. In the coast of my country, squid mansoni is an economical cephalopod with abundant resources and good yield. The needleless squid is full of treasures, and has high edible and medicinal value. It was one of the "four sea products" in my country in the 1970s and 1980s. Its annual output can reach tens of thousands of tons, of which...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2563/107C12Q2545/114
Inventor 高天翔陈治
Owner ZHEJIANG OCEAN UNIV
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