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Method for quickly and high-efficiently detecting sepia esculenta hoyle population

A golden squid, high-efficiency technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of harming target species, high cost, time-consuming and labor-intensive, etc., to achieve improved efficiency and accuracy, rapid and efficient detection , the effect of reducing the cost of testing

Inactive Publication Date: 2018-05-04
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 golden squid population distribution, on-site survey methods are mainly used, such as fishing with nets, which is time-consuming, labor-intensive, high in cost, low in catch rate, and may harm the target species or destroy the ecosystem of the survey site. and other shortcomings, but there is no method for quickly and effectively detecting the population of golden squid without damaging the environment in the prior art. Method for fast and efficient detection of golden squid populations for detection of squid populations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A method for fast and efficient detection of golden squid populations, the method for fast and efficient detection of golden squid populations 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 using specially-made primers and probes. The special primers were the upstream primer SEQ ID NO.1: 5'-AGCAGGAACAGGATGAAC-3' and the downstream primer SEQ ID NO.2:5 '-GCCAAATCGACAGAAGGT-3', the probe sequence is SEQ ID NO.3: 5'-FAM-CTACCCTCCTTTATCAAGTAACCTCTCAC-TAMRA-3';

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

Embodiment 2

[0037] A method for fast and efficient detection of golden squid populations, the method for fast and efficient detection of golden squid populations 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 is carried out with special primers and probes, the special primers are the upstream primer SEQ ID NO.1:5'-AGCAGGAACAGGATGAAC-3' and the downstream primer SEQ ID NO.2:5 '-GCCAAATCGACAGAAGGT-3', the probe sequence is SEQ ID NO.3: 5'-FAM-CTACCCTCCTTTATCAAGTAACCTCTCAC-TAMRA-3';

[0041] The composition of the qPCR amplification reaction system is: 10 μL of PremixExTaq, 0.4...

Embodiment 3

[0045] A method for fast and efficient detection of golden squid populations, the method for fast and efficient detection of golden squid populations 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 is carried out with special primers and probes, the special primers are the upstream primer SEQ ID NO.1:5'-AGCAGGAACAGGATGAAC-3' and the downstream primer SEQ ID NO.2:5 '-GCCAAATCGACAGAAGGT-3', the probe sequence is SEQ ID NO.3: 5'-FAM-CTACCCTCCTTTATCAAGTAACCTCTCAC-TAMRA-3';

[0049] The composition of the qPCR amplification reaction system is: 10 μL ...

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Abstract

The invention relates to a method for quickly and high-efficiently detecting a sepia esculenta hoyle population. The method can be used for detecting the sepia esculenta hoyle population through collecting environment DNA (Deoxyribonucleic Acid) without damaging an environment and a species population. The collected eDNA samples are sampled from soil, silt, water and the like, so that an environment burden cannot be caused on a detecting point, and an adverse effect cannot be caused on a species survival environment; a special primer and a special probe are utilized for accurately, quickly andhigh-efficiently detecting DNA of sepia esculenta hoyle, so that the detection efficiency and the accuracy are improved, and the detection cost is reduced; compared with a field investigation methodutilized by traditional sepia esculenta hoyle population distribution detection, the method has the advantages of saving both time and labor, low cost, high efficiency, and no harm to target species or damaging an ecological system of a survey spot.

Description

technical field [0001] The invention relates to a method for detecting golden squid populations, in particular to a method for rapidly and efficiently detecting golden squid populations by using environmental DNA to detect golden squid populations without damaging the environment and species populations. Background technique [0002] Sepia esculenta (Hoyle 1885), also known as cuttlefish and mullet, belongs to mollusca phylum Mollusca, cephalopod class Cephalopoda, sheath subclass Coleoidea, squid order Sepioidea, squid family Sepiidea, squid genus Sepia, distributed in Bohai Sea, Yellow Sea, East China Sea , the South China Sea, the Japanese Archipelago, and the Philippine Islands. The golden squid is one of the most important economic species in the world squid family, and its annual output ranks second in the world squid species. It is the squid with the largest economic value in the northern seas of my country. One of the four traditional fisheries, it is an important fi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2531/113C12Q2563/107
Inventor 高天翔陈治
Owner ZHEJIANG OCEAN UNIV
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