Molecular marker method for identifying fugu rubripes parentage as well as microsatellite and kit used for molecular marker method

A parent-child relationship and microsatellite technology, applied in the field of molecular markers, can solve the problems of slow growth of fish fry and high cost of individual electronic markers

Inactive Publication Date: 2014-06-18
CHINESE ACAD OF FISHERY SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, it is also possible to electronically mark individuals and then mix them in a large cement pool to reduce the impact of the environment on different families. The family is raised in an independent water tank for 4-5 months, and the electronic marking can only

Method used

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  • Molecular marker method for identifying fugu rubripes parentage as well as microsatellite and kit used for molecular marker method
  • Molecular marker method for identifying fugu rubripes parentage as well as microsatellite and kit used for molecular marker method
  • Molecular marker method for identifying fugu rubripes parentage as well as microsatellite and kit used for molecular marker method

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Embodiment 1

[0042] The present invention is a molecular marker method for identifying the pure parent-child relationship of Redfin Oriental, which adopts the following technical steps:

[0043] There are a total of 272 fish in 5 families of Hongfin Dongfangchun, including 5 pairs of parents, collected from the Beidaihe Central Experimental Station of the Chinese Academy of Fishery Sciences. The ventral fin rays of each fish were collected, and the genomic DNA was extracted with the TIANGEN Marine Animal Genomic DNA Extraction Kit. Using the extracted genomic DNA as a template, perform PCR amplification with the microsatellite markers with the top 5 exclusion rates. The total volume of the PCR reaction is 25 μL, including: 10×Buffer2.5 μL, Mg 2+ (25mmol / L) 1μL, dNTPs (2mmol / L each) 1μL, forward and reverse primers (10μrnol / L) 1μL each, template DNA 1μL (50ng / μL), Taq DNA polymerase 1U, add appropriate amount of ddH 2 O. The PCR reaction program was: pre-denaturation at 94°C for 3 min, 30...

Embodiment 2

[0049] The present invention is a molecular marker method for identifying the pure parent-child relationship of Redfin Oriental, which adopts the following technical steps:

[0050]There are 313 fish in 7 families of Hongfin Dongfangchun, including 7 pairs of parents, collected from the Beidaihe Central Experimental Station of the Chinese Academy of Fishery Sciences. The ventral fin rays of each fish were collected, and the genomic DNA was extracted with the TIANGEN Marine Animal Genomic DNA Extraction Kit. Using the extracted genomic DNA as a template, perform PCR amplification with the microsatellite markers with the top 5 exclusion rates. The total volume of the PCR reaction is 25 μL, including: 10×Buffer2.5 μL, Mg 2+ (25mmol / L) 1μL, dNTPs (2mmol / L each) 1μL, forward and reverse primers (10μmol / L) 1μL each, template DNA 1μL (50ng / μL), Taq DNA polymerase 1U, add appropriate amount of ddH 2 O. The PCR reaction program was: pre-denaturation at 94°C for 3 min, 30 cycles (dena...

Embodiment 3

[0057] The present invention is a molecular marker method for identifying the pure parent-child relationship of Redfin Oriental, which adopts the following technical steps:

[0058] There are 359 fish in 8 families of Hongfin Dongfangchun, including 8 pairs of parents, collected from the Beidaihe Central Experimental Station of the Chinese Academy of Fishery Sciences. The ventral fin rays of each fish were collected, and the genomic DNA was extracted with the TIANGEN Marine Animal Genomic DNA Extraction Kit. Using the extracted genomic DNA as a template, perform PCR amplification with the microsatellite markers with the top 5 exclusion rates. The total volume of the PCR reaction is 25 μL, including: 10×Buffer2.5 μL, Mg 2+ (25mmol / L) 1μL, dNTPs (2mmol / L each) 1μL, forward and reverse primers (10μmol / L) 1μL each, template DNA 1μL (50ng / μL), Taq DNA polymerase 1U, add appropriate amount of ddH 2 O. The PCR reaction program was: pre-denaturation at 94°C for 3 min, 30 cycles (den...

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Abstract

The invention provides a method for identifying fugu rubripes parentage by using a molecular marker method as well as microsatellite primers and a kit used for the molecular marker method. Selected 44 microsatellite primers are stable in pure PCR amplification reaction, clear in amplified fragment and high in polymorphism. By using the technology, different families of mixed fugu rubripes individuals can be distinguished, so that different families of the fugu rubripes individuals can be quickly and accurately identified, and the technology has great application value for breed conservation, propagation and breeding selection of fugu rubripes.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and relates to a method for identifying the pure parent-child relationship of red fin orientals by using molecular markers, as well as a microsatellite primer and a kit. Background technique [0002] Takifugu rubripes belongs to Osteichthyes, Tetraodontiformes, Tetraodontoidei, Tetraodntoidea, Tetraodontidae, Takifugu, Commonly known as "blowfish", it is mainly produced in the Bohai Sea, the Yellow Sea and the East China Sea in my country, and it is mainly distributed in the coast of Japan and the Korean Peninsula abroad. The red fin Dongfang pure meat is tender, delicious, high in protein and low in fat, rich in vitamins and trace elements, known as the "king of fish". Although Redfin Dongfangchun has relatively comprehensive and excellent traits, there are no varieties in the actual breeding process at this stage, and there are relatively few selective breeding studies on Redfin Dongf...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2531/113
Inventor 刘永新周勤张红涛
Owner CHINESE ACAD OF FISHERY SCI
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