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Microsatellite molecular marker sites of big pacific salmons, and polymorphic primers and application thereof

A technology of molecular marker and salmon, applied in the field of molecular biology DNA molecular marker, can solve problems such as resource reduction, and achieve the effect of accurately evaluating the effect of salmon proliferation and release

Active Publication Date: 2020-02-07
FISHERIES SCI RES INST OF JILIN PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the deterioration of its living environment in recent years, its resources have decreased sharply, and it is now in an endangered state. It is urgent to vigorously carry out the protection of aquatic germplasm resources.

Method used

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  • Microsatellite molecular marker sites of big pacific salmons, and polymorphic primers and application thereof
  • Microsatellite molecular marker sites of big pacific salmons, and polymorphic primers and application thereof
  • Microsatellite molecular marker sites of big pacific salmons, and polymorphic primers and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Screening of Microsatellite Molecular Markers

[0028] According to the SSR analysis carried out in the constructed salmon fin ray tissue genomic library, the present invention obtained 50 microsatellite molecular marker sites by comparison with the genomic library, and designed 50 pairs of screening primer sequences according to the relevant sites. Salmon 16 individuals were verified.

Embodiment 2

[0029] The verification of embodiment 2 microsatellite primers

[0030] Using the salmon genome as a template, 50 pairs of primer sequences were used for temperature gradient PCR to find out the optimal annealing temperature, and the PCR amplification results were detected by 1% agarose gel electrophoresis.

[0031] PCR system: 2×Taq PCR mix 12.5 μL; primers (5uM) 1 μL; DNA template 1 μL; ddH 2 0 to 25 μL. PCR program: 94°C for 5min, (94°C for 30s, X°C for 30s, 72°C for 30s)×35 cycle, 72°C for 5min. Screen out the primer pairs with the correct target band.

Embodiment 3

[0032] Example 3 microsatellite primer polymorphism verification

[0033] The primers with bands obtained in Example 2 were synthesized with fluorescent primers, and the 5' of one of the primers was marked with FAM / HEX / Tamra. PCR amplification detection, the primer amplification effect picture is as follows Figure 1-2 As shown, it can be seen from the figure that using this PCR system to amplify can obtain a very good primer amplification effect.

[0034] PCR system: 10×PCR buffer 1μL; primers (10mM) 0.4μL; dNTP 0.6μL; DNA template 1μL; Taq enzyme 0.5μL; ddH 2 0 to 10 μL. PCR program: 94°C for 2min, (94°C for 30s, 60°C to 50°CΔ1°C for 30s, 72°C for 30s)×50 cycle, (94°C for 30s, 50°C for 30s, 72°C for 30s)×30 cycle, 72°C for 5min. The product was analyzed with ABI3730xl automatic DNA sequencer. Software Cervus3.0 carries out genetic index (allele (na: Observed number of alleles), observed heterozygosity (H O : Observed heterozygosity), expected heterozygosity (H E : Expe...

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Abstract

The invention discloses microsatellite molecular marker sites of big pacific salmons as well as polymorphic primers and application thereof. The invention discloses microsatellite molecular marker sites of big pacific salmons as well as polymorphic primers and application thereof. According to the invention, 10 microsatellite loci are screened from genomic DNA of big pacific salmons; and specificprimers are designed on flanking sequences at both ends of the microsatellite loci according to the 10 screened microsatellite loci. Polymorphism and stability are shown by amplification results usingthe primers, so that the microsatellite loci are suitable for the fields of genetic relationship identification, genetic diversity detection of population and molecule-assisted breeding of the big pacific salmons.

Description

technical field [0001] The invention belongs to the technical field of molecular biology DNA molecular markers, in particular to the development technology of salmon microsatellite molecular markers. Background technique [0002] The salmon Oncorhynchus keta (Walbaum) belongs to the family Salmonidae and belongs to the Pacific salmon genus. It is anadromous, that is, it lives in the river, grows in the sea and returns to the river of origin to lay eggs and reproduce. Salmon is a valuable economic fish in the world, and its roe is also a famous aquatic product with high nutritional value. It is famous for its delicious meat and rich nutrition, and has always been regarded as a valuable fish. Salmon is also a famous cold-water fish in the Tumen River Basin in Changbai Mountain, Jilin Province. However, due to the deterioration of its living environment in recent years, its resources have decreased sharply, and it is now in an endangered state. It is urgent to vigorously carry...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 郑伟韩叶闫春梅万继武李忠强马驰刘慧吉王战蔚
Owner FISHERIES SCI RES INST OF JILIN PROVINCE
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