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Molecular marking method for discriminating different family of Cyprinus carpiovar jian

A molecular marker and pedigree technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of slow growth of fry, large fish ponds, and high cost of individual identification, and achieve stable PCR amplification reaction and polymorphism. High performance and clear amplified fragments

Inactive Publication Date: 2013-03-27
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the comparison test of breeding, different families are usually placed in different ponds or isolated in different cages for breeding, which requires a lot of fish ponds and equipment. In addition, it is often difficult to achieve due to different pond water quality environments. One to one, it is easy to cause errors in the results of breeding comparison experiments
At present, it is also possible to carry out breeding comparison experiments in the same water area after individual identification (such as PIT marking) to reduce the impact of the environment on different families. However, this method also has certain drawbacks: it is still necessary to identify different The offspring of the family are kept in isolation for 2-3 months in small pools or net cages, and can only be identified when the weight grows to about 30g, and the breeding during this period will also have a great impact on the growth of fry --- in small pools or net cages The fish fry cultured in isolated boxes grow much slower than those directly cultured in large ponds
Also, the cost of individual marking is relatively high. Taking PIT marking as an example, the price of one PIT marking is about 20 yuan

Method used

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  • Molecular marking method for discriminating different family of Cyprinus carpiovar jian
  • Molecular marking method for discriminating different family of Cyprinus carpiovar jian
  • Molecular marking method for discriminating different family of Cyprinus carpiovar jian

Examples

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Effect test

Embodiment 1

[0032] A kind of molecular marker method for distinguishing different families of Jian carp of the present invention adopts the following technical steps:

[0033] There are 28 fish in 3 families of Jian carp, including 3 pairs of parents, and 6-8 offspring in each family. They were collected from Yixing Fishery, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences. 0.5 mL of blood was collected from the tail vein of each fish, and 30 μL of blood cells were taken to extract genomic DNA. Using the extracted genomic DNA as a template, 12 pairs of primers were used for PCR amplification respectively. The total volume of PCR reaction was 10 μL, which contained 1.0 μL of 10× reaction buffer, Mg2+ 2 mmol / L, dNTP 200 μmol / L, upstream and downstream 0.2 μmol / L of each primer, 0.3 U of Taq enzyme, 50 ng to 80 ng of DNA, and make up the volume with sterilized double distilled water. The PCR reaction conditions were: 94°C for 3 min; 94°C for 30 s, annealing for 30 s...

Embodiment 2

[0037] A kind of molecular marker method for distinguishing different families of Jian carp of the present invention adopts the following technical steps:

[0038] A total of 50 fish in 5 families of Jian carp, including 5 pairs of parents, and 8 offspring in each family, were collected from Yixing Fishery, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences. 0.5 mL of blood was collected from the tail vein of each fish, and 30 μL of blood cells were taken to extract genomic DNA. Using the extracted genomic DNA as a template, 12 pairs of primers were used for PCR amplification respectively. The total volume of PCR reaction was 10 μL, which contained 1.0 μL of 10× reaction buffer, Mg2+ 2 mmol / L, dNTP 200 μmol / L, upstream and downstream 0.2 μmol / L of each primer, 0.3 U of Taq enzyme, 50 ng to 80 ng of DNA, and make up the volume with sterilized double distilled water. The PCR reaction conditions were: 94°C for 3 min; 94°C for 30 s, annealing for 30 s, 72°C...

Embodiment 3

[0042] A kind of molecular marker method for distinguishing different families of Jian carp of the present invention adopts the following technical steps:

[0043]There are 58 fish in 6 families of Jian carp, including 6 pairs of parents, and 6-8 offspring in each family. They were collected from Yixing Fishery, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences. 0.5 mL of blood was collected from the tail vein of each fish, and 30 μL of blood cells were taken to extract genomic DNA. Using the extracted genomic DNA as a template, 12 pairs of primers were used for PCR amplification respectively. The total volume of PCR reaction was 10 μL, which contained 1.0 μL of 10× reaction buffer, Mg2+ 2 mmol / L, dNTP 200 μmol / L, upstream and downstream 0.2 μmol / L of each primer, 0.3 U of Taq enzyme, 50 ng to 80 ng of DNA, and make up the volume with sterilized double distilled water. The PCR reaction conditions were: 94°C for 3 min; 94°C for 30 s, annealing for 30 s,...

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Abstract

The invention relates to a molecular marking method for discriminating different families of Cyprinus carpiovar jian. The method is characterized by comprising steps of: selecting 12 pairs of specific microsatellite marker primers; sampling blood or pterygiophore from a Cyprinus carpiovar jian individual and parents thereof of different families; extracting genome DNA; carrying out PCR amplification on the extracted genome DNA; carrying out electrophoretic separation on the PCR products; carrying out digitalized processing on the PCR electrophoresis detection results; analyzing the results by a piece of heredity analysis software to obtain heredity distance among each individual and construct a cladogram; and discriminating individuals from different families according to a cluster result of heredity distances among individuals. The invention can discriminate different families of Cyprinus carpiovar jian rapidly and effectively and provide assistance for seed selection and breeding setting of Cyprinus carpiovar jian.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and relates to a molecular marker method for identifying different families of Jian carp, which is used to identify different families of Jian carp. Background technique [0002] Carp is one of the most important freshwater aquaculture species in my country, Jian carp ( Cyprinus carplo vat. jian ) is a carp variety with stable genetic traits successfully bred by the comprehensive breeding technology of family selection, multi-line hybridization and gynogenesis. High adaptability and strong resistance to adverse conditions. Although Jian carp has relatively comprehensive and excellent traits, as an artificially bred variety, it still needs continuous selection, conservation and genetic protection, and it is very necessary to breed new strains with better traits through genetic improvement. Multi-line selection is a method widely used in aquatic breeding. The method of multi-line sele...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 李建林俞菊华唐永凯李红霞董在杰
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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