A rapid identification method for the hybrids of the high-body crocodile and the veschia crocodile

A technology of crocodile and crocodile, applied in biochemical equipment and methods, measurement/inspection of microorganisms, etc., can solve the problem of mixed germplasm, unfavorable breeding of new species with salt-alkali tolerance, large water surface breeding, juvenile The phenotype determination characteristics of fish and adult fish are not obvious, etc., to achieve the effect of less non-specific fragments, clear amplification bands, accurate and rapid identification

Active Publication Date: 2021-10-15
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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Problems solved by technology

[0003] In view of the lack of obvious phenotypic characteristics of the parent and hybrid seedlings and adult fish, it is easy to cause germplasm mixing, which is not conducive to the demonstration and application of new saline-alkali-tolerant varieties in saline-alkali pond culture and large-water surface breeding.

Method used

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  • A rapid identification method for the hybrids of the high-body crocodile and the veschia crocodile
  • A rapid identification method for the hybrids of the high-body crocodile and the veschia crocodile

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specific Embodiment approach 1

[0022] Specific implementation mode one: the present embodiment carries out the method for rapid identification of the high body crocodile and the Washi crocodile hybrid:

[0023] 1. Extraction of genomic DNA from the sample to be tested;

[0024] 2. SSR-PCR reaction;

[0025] 3. Gel electrophoresis;

[0026] Four, identification of hybrid species: 2 allele bands appear, wherein one allele band size is 95bp or 107bp, and the other allele band size is 101bp, then it is determined that the sample is a high body crocodile and Varsity crocodile hybrids;

[0027] Wherein, in step two SSR-PCR reaction, specific SSR labeling primers are Primer1-F and Primer1-R,

[0028] Primer1-F: 5'-TGATGGTGGTGGTGGAGATG

[0029] Primer1-R: 5'-TGCACAGTTCCACTGAGTGT.

specific Embodiment approach 2

[0030] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the SSR-PCR reaction system in step 2 is 15 μL, which consists of 10.8 μL self-made mixed buffer, 0.5 μL forward detection primer, 0.5 μL reverse detection primer, 2 μL Sample DNA, 0.2 μL Taq DNA polymerase with an activity of 1U and 1 μL deionized sterile water; 0.5 μL forward identification primer contains forward identification primer Primer1-F 10mmol / L, 0.5 μL reverse identification primer Contain reverse identification primer Primer1-R10mmol / L; 10.8μL self-made mixed buffer contains 50mmol / L KCl, 10mmol / L Tris-HCl, 0.1% volume TritonX-100, 1.5mmol / L MgCl 2 , 0.1% volume of NP-40, 0.01% volume of gelatin and 200 μmol / L of four kinds of dNTP mixture. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0031] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that 2 μL of sample DNA in the SSR-PCR reaction system in step 2 contains 50 ng of sample DNA. Other steps and parameters are the same as those in Embodiment 1 or 2.

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Abstract

The invention discloses a method for quickly identifying the hybrid species of the high-body crocodile and the Washii crocodile, relating to a method for quickly identifying the hybrid fish. The invention aims at distinguishing the parents and hybrids of the high-body crocodile and the crocodile veschii, especially in the early stage of the juvenile fry. Rapid identification methods: 1. Extraction of genomic DNA from the sample to be tested; 2. SSR-PCR reaction; 3. Gel electrophoresis; 4. Hybrid identification: 2 allele bands appear, and the size of one allele band is 95bp or 107bp, and the other allele band size is 101bp, then it is determined that the sample is a hybrid of the high-body crocodile and the Waschner crocodile. The invention has the advantages of simple and convenient operation, good repeatability, good versatility, is not affected by environmental factors, and can accurately and quickly identify the authenticity of the hybrid progeny.

Description

technical field [0001] The invention relates to a method for quickly identifying hybrid fish. Background technique [0002] Leuciscus idus and Leuciscus waleckii are two different species belonging to Cyprinidae, Leuciscus subfamily, and Leuciscus genus. The high-body crocodile is mainly distributed in the Irtysh River system in Xinjiang in my country. It is wide and grows fast; the crocodile is mainly distributed in Heilongjiang, Yellow River, Liaohe and inland saline-alkaline lakes. It has strong salt-alkali tolerance. It can withstand the harsh water environment with an alkalinity of more than 50mmol / L and a pH of 9.6 in Dali Lake, Inner Mongolia. There are about 960 million mu of low-lying saline-alkali waters in my country, which have the characteristics of high alkalinity, high pH, ​​and complex ion composition. The utilization rate is less than 2%, and most of them are in a state of abandonment. Practice has proved that "using fish to control alkali" is an effective ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6858
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 常玉梅梁利群赵雪飞孙博张立民
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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