Standard black sea bream DNA bar code detecting sequence and application thereof

A standard detection and barcoding technology, applied in recombinant DNA technology, DNA/RNA fragments, introduction of foreign genetic material using vectors, etc., can solve problems such as lack of data, achieve high repeatability, shorten identification time, and good stability.

Inactive Publication Date: 2017-08-11
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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  • Standard black sea bream DNA bar code detecting sequence and application thereof
  • Standard black sea bream DNA bar code detecting sequence and application thereof
  • Standard black sea bream DNA bar code detecting sequence and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Collection, storage and processing of samples to be tested

[0043] Part of the muscle tissue of the black sea bream was cut from the soaked specimen, stored in 95% alcohol and stored in a -20°C refrigerator.

[0044] 2. DNA extraction

[0045] The DNA in the samples was extracted by the phenol-chloroform method and stored at -20°C for future use.

[0046] 3. Synthesis of PCR primers

[0047] This method selects 2 pairs of universal primers:

[0048] FishF2_t1:

[0049] 5'-TGTAAAACGACGGCCAGTCGACTAATCATAAAGATATCGGCAC-3' (SEQ ID NO: 2);

[0050] FishR2_t1:

[0051] 5'-CAGGAAACAGCTATGACACTTCAGGGTGACCGAAGAATCAGAA-3' (SEQ ID NO: 3);

[0052] VF2_t1:

[0053] 5'-TGTAAAACGACGGCCAGTCAACCAACCACAAAGACATTGGCAC-3' (SEQ ID NO: 4);

[0054] FR1d_t1:

[0055] 5'-CAGGAAACAGCTATGACACCTCAGGGTGTCCGAARAAYCARAA-3' (SEQ ID NO: 5). Wherein, the base at R is A or G, the base at Y is C or T, and the establishment of R or Y is used to amplify unknown bases.

[0056] 4. PCR reaction...

Embodiment 2

[0069] Two groups of muscle tissue samples were selected from fish tissue samples (incomplete, unable to carry out biological identification), and preserved in 95% alcohol. The selected muscle tissue belonged to two groups of different species of fish, one group was black sea bream (5 samples), and the other group was other species (5 samples), and the sampler recorded and numbered the species according to the specific information. The experiment operator conducted the experiment without knowing the species of the two groups of samples. Adopt the same experimental method as Example 1 to carry out the experiment, PCR product sequencing results show that 5 samples of one group are tissue samples of black sea bream, and 5 samples of another group are tissue samples of non-black sea bream, and the experimental results are the same as sampling The records are exactly the same.

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Abstract

The invention discloses a standard black sea bream DNA bar code detecting sequence and application thereof. The sequence is shown as SEQ ID NO:1. By adopting a black sea bream molecule identification method, a reliable DNA identification technology adopted for individuals which are incomplete or cannot accurately identified in specimens and complete specimens is achieved, and the technical defect that in the prior art, incomplete individuals cannot be identified and complete individuals can be identified by needing specialized persons very familiar with the morphology of black sea breams is overcome. The black sea breams can be rapidly and accurately identified compared with a traditional morphological identification method.

Description

technical field [0001] The invention relates to the technical field of species identification, and relates to a black sea bream DNA barcode standard detection sequence and its application in species identification. Background technique [0002] Black sea bream (scientific name: Acanthopagrus schlegelii), also known as black sea bream, commonly known as Heijiaji, black fin, etc., is a fish belonging to the genus Acanthopagrus schlegelii of the order Perciformes, and is the most popular economic fish in the coast of my country. It is one of the species and is also an important mariculture species. Mainly distributed in the Western Pacific region, including the East China Sea, Hong Kong, Japan and other waters. Black sea bream is a wide-temperature and wide-halt fish, and the maximum individual length can reach 45 cm. Generally, the growth rate before the 4th age is relatively fast. At present, the species identification of black sea bream is limited to morphological identific...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/63
Inventor 游欣欣张凯马兴宇
Owner ZHENJIANG HUADA DETECTION CO LTD
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