Silurus asotus DNA barcoding standard detection sequence and application thereof

A standard detection and barcode technology, applied in the direction of recombinant DNA technology, DNA / RNA fragments, applications, etc., can solve the problem of no data, achieve high repeatability, good stability, and shorten the identification time

Inactive Publication Date: 2017-08-18
ZHENJIANG HUADA DETECTION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the endemism of many fishes in

Method used

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  • Silurus asotus DNA barcoding standard detection sequence and application thereof
  • Silurus asotus DNA barcoding standard detection sequence and application thereof
  • Silurus asotus DNA barcoding standard detection sequence and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0034] Example 1:

[0035] 1. Collection, storage and processing of samples to be tested

[0036] Part of the muscle tissue of the catfish was cut from the immersed specimen and stored in 95% alcohol in a refrigerator at -20°C.

[0037] 2. DNA extraction

[0038] The DNA in the sample is extracted by the kit method of the biological company and stored at -20°C for later use.

[0039] 3. PCR primer synthesis

[0040] This method selects 1 pair of primers:

[0041] NYCOI-F2: 5'-CCTCCTTCCTACTACTAT-3' (SEQ ID NO: 2);

[0042] NYCOI-R2: 5'-CCAAGAATGGAGGACACC-3' (SEQ ID NO: 3);

[0043] 4. PCR reaction system (25μL)

[0044]

[0045] 5. PCR reaction program

[0046]

[0047] After the reaction is over, turn on the PCR machine, take out the amplified sample, and store it at 4°C

[0048] 6. PCR product detection

[0049] Take 2μl of PCR product, use 1.5% agarose gel electrophoresis (120V, 200mA, 25min), stain with genegreen dye. Gel imaging system detection, electrophoresis pattern such as figure 2 ...

Example Embodiment

[0054] Example 2:

[0055] Two groups of muscle tissue samples were selected from fish tissue samples (incomplete and unable to conduct biological identification) and stored in 95% alcohol. The selected muscle tissues belong to two groups of fish of different species, one of which is catfish (10 samples), and the other group is other species (10 samples). The sampler will record and number the species according to specific information. The operator conducts the experiment without knowing the species belonging to the two groups of samples. The experiment was carried out using the same experimental method as in Example 1. The PCR product sequencing results showed that 10 samples in one group were catfish tissue samples, and the other 10 samples were non-catfish tissue samples. The experimental results were recorded with the sampler The information is exactly the same.

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Abstract

The invention discloses a silurus asotus DNA barcoding standard detection sequence and application thereof. The sequence is shown as SEQ ID NO:1. The silurus asotus molecular identification method provided by the invention realizes the reliable DNA molecular identification technology that solves the difficulties in identification of silurus asotus, silurus soldatovi Meridionalis Chen and other silurus asotus species, overcomes the technical defects of need for professionals very familiar with the silurus asotus morphology during complete individual identification in the prior art, and compared with the traditional morphological identification method, can rapidly and accurately identify silurus asotus.

Description

technical field [0001] The invention relates to the technical field of species identification, in particular to a catfish DNA barcode standard detection sequence and its application in species identification. Background technique [0002] Catfish (scientific name: Silurus asotus) belongs to the genus Cyprinidae of the subclass Cypriniformes of the ray-finned fishes, and is a unique species in China. Similar species of catfish are distributed almost all over the world, and most species live in fresh water such as rivers or rivers, but some species live in oceans. At present, the species identification of catfish is limited to morphological identification, and there are few professional identification personnel. Due to the destruction of spawning grounds due to the construction of dams in the upper reaches of the Yangtze River, and the invasion of alien species and water pollution, the living environment of catfish has been severely damaged, so the population of catfish is de...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/12C12N15/11
Inventor 石琼游欣欣崔海月蒋寿佳耿小燕
Owner ZHENJIANG HUADA DETECTION CO LTD
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