Molecular biological method for rapidly identifying two leuciscus waleckii species distributed in same area of Sinkiang

A technology of molecular biology and crocodile, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem of lack of universal primers for amplifying fish mitochondrial 12SrRNA gene, destruction of high-body crocodile resources, research Difficulties and other problems, to achieve the effect of improving accuracy and stability, increasing success, and high credibility

Inactive Publication Date: 2019-01-22
ZHEJIANG OCEAN UNIV
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Problems solved by technology

The difference in the morphological classification key table between the two is only reflected in the number of lateral line scales (the former is 46-52, the latter is 56-61) and the ratio of body length to body height (the former body length / body height is about 3.7-4.5, and the latter body length / body height about 3.2-3.7), especially in the juvenile stage, it is difficult to distinguish with the naked eye, and it is easy to cause accidental catch and further damage the resources of the tall crocodile
At present, the use of mitochondrial 12S rRNA gene sequences as molecular markers has been widely used in the fields of fish molecular population genetics and phylogenetics. certain difficulty

Method used

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  • Molecular biological method for rapidly identifying two leuciscus waleckii species distributed in same area of Sinkiang

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Embodiment 1

[0024] A molecular biology method for rapid identification of two sympatrically distributed yalo fish in Xinjiang, comprising the following steps:

[0025] 1) Using the improved phenol-chloroform method to extract the genomic DNA of the Yaluo fish tissue respectively;

[0026] 2) designing primers for the mitochondrial 12S rRNA gene;

[0027] 3) using the DNA of the two fishes as templates for PCR amplification;

[0028] 4) PCR products were detected by 1% agarose gel electrophoresis, and PCR products with clear bands were selected for sequencing;

[0029] 5) sequence alignment;

[0030] The tissue extract for the modified phenol-chloroform method contains 10 mM Tris-Cl, 0.1 M EDTA, 0.32 mM DTE, 0.04 mMBJ-BR, and 0.5% SDS; the pH of the tissue extract is 7.8. Each component in the tissue extract can play a beneficial role, improve the digestion effect of the extract on the caudal fin tissue, increase the amount of protein released, and at the same time, it can promote its a...

Embodiment 2

[0042] A molecular biology method for rapid identification of two sympatrically distributed yalo fish in Xinjiang, comprising the following steps:

[0043] 1) The two samples of Yaluo fish were collected from the Irtysh River Basin in Xinjiang. After the morphological and anatomical identifications were confirmed, the fresh caudal fin tissues were cut and fixed in absolute ethanol, and brought back to the laboratory and placed in- Stored in a refrigerator at 20°C, the genomic DNA of the crocodile tissue was extracted by the modified phenol-chloroform method; %SDS; the pH of the tissue extract is 7.8;

[0044] 2) Design primers for the mitochondrial 12S rRNA gene: download the rDNA sequence of the crocodile fish from the Genbank database, perform multiple alignments on these sequences, and find relatively conserved sequences and highly variable sequences at the 3' end and 5' end, respectively. region, and design primers to amplify the mitochondrial 12S rRNA gene on the 12S rRN...

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Abstract

The invention discloses a molecular biological method for rapidly identifying two leuciscus waleckii species distributed in the same area of Sinkiang. The molecular biological method comprises the following steps: (1) extracting genomes of leuciscus waleckii tissue by adopting an improved phenol-chloroform method; (2) designing primers of mitochondria 12S rRNA genes; (3) by adopting DNAs of the two fish species as templates, carrying out PCR amplification; (4) carrying out 1% agarose gel electrophoresis detection on the PCR products, selecting the PCR products with clear stripes, and carryingout sequencing; and (5) carrying out sequence contrast. The molecular biological method has the beneficial effects that the molecular biological method is scientific and effective, is simple and rapid, is accurate and feasible, is strong in specificity, and is high in reliability, leuciscus waleckii does not need to be killed, amplification can be carried out on the two species at once, and thus the operation is efficient, rapid and convenient.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a molecular biology method for rapidly identifying two cymbidiums distributed in the same region in Xinjiang. technical background [0002] Baikal croaker (Leuciscus baicalensis), also known as small white fish, small white stripes. Leuciscus idus, also known as medium white fish, both belong to Cyprinidae, Cyprinidae, Leuciscus subfamily, and Leuciscus genus. my country is only distributed in Irtysh River and Ulun, Xinjiang The ancient river system is an important indigenous economic fish in this basin. Among them, due to its fresh and tender meat and high nutritional value, the high-tire crocodile has been under too much fishing pressure in recent years, and its resources have declined sharply. In 2006, it was listed as the second-level protected fish in the autonomous region, and it is a key protected aquatic wild animal in the autonomous region. The difference in t...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2531/113
Inventor 杨天燕孟玮高天翔
Owner ZHEJIANG OCEAN UNIV
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