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Scophthalmus maximus T170G single nucleotide polymorphic marking detection method

A single nucleotide polymorphism and detection method technology is applied in the field of detection of turbot T170G single nucleotide polymorphism markers, which can solve the problems of cumbersome operation and achieve simple operation, error reduction, and high-throughput closed-tube The effect of the operation

Inactive Publication Date: 2013-09-11
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

However, when they analyze EST sequences, the software they use is CAP3 and QualitySNP. The operation of this software requires the Linux operating system, and the operation is relatively cumbersome.

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  • Scophthalmus maximus T170G single nucleotide polymorphic marking detection method
  • Scophthalmus maximus T170G single nucleotide polymorphic marking detection method
  • Scophthalmus maximus T170G single nucleotide polymorphic marking detection method

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Embodiment Construction

[0023] A method for detecting single nucleotide polymorphism markers of turbot T170G, comprising 1), extraction of genomic DNA; 2), screening of candidate SNPs; 3), design of primers and probes; 4), asymmetric PCR amplification 5), probe hybridization; 6), data acquisition; 7), genotyping, the specific steps are as follows:

[0024] 1. Genomic DNA extraction:

[0025]The samples in this experiment came from 42 individuals of 10 different trait groups in 8 families of Yantai Tianyuan Aquatic Products Co., Ltd., and the DNA was extracted by using the marine animal tissue genomic DNA extraction kit (spin column type, TIANGEN). a. Take 20 mg of fresh muscle tissue, put it into a centrifuge tube filled with 200 μL GA buffer, and vortex for 15 seconds. b. Add 20 μL proteinase K (20mg / ml) solution, vortex to mix, briefly centrifuge to remove the water droplets on the inner wall of the tube cap, place at 56°C until the tissue is completely dissolved, and briefly centrifuge to remove ...

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Abstract

The invention relates to a scophthalmus maximus T170G single nucleotide polymorphic marking detection method. The method comprises the following steps: extracting scophthalmus maximus genome DNA and diluting for later use; analyzing the sequences of EST, screening the sequence containing a candidate SNP locus, designing a specific primer at the two ends and designing a non-marking probe with the closed 3' end in front of and behind the locus (comprising the locus); performing asymmetric PCR amplification on the scophthalmus maximus group genome DNA by using the primer; hybridizing the amplification product with the non-marking probe with the closed 3' end; and placing the hybrid product on a LightScanner, and detecting and analyzing the melting curve to obtain the genetic polymorphism map of the scophthalmus maximus. The method is applicable to detection technologies of scophthalmus maximus genetic marking, genealogy authentication, genetic linkage map construction and the like. According to the method which is convenient, quick and accurate, the scophthalmus maximus T170GSNP marked genetic variation map can be obtained quickly; and the genotype of each individual of the scophthalmus maximus can be detected intuitively.

Description

Technical field: [0001] The invention belongs to molecular genetic marker research technology, in particular to a method for detecting turbot T170G single nucleotide polymorphism marker (single nucleotide polymorphism, SNP). Background technique: [0002] Single nucleotide polymorphism (SNP) molecular marker is the most abundant mutation type in the genome, because of its large number, high coverage density, rich loci, almost distributed in the entire genome, with short fragments, easy amplification, and genetic stability Strong and other characteristics; easy to high-throughput, automated analysis, easy to study genetic mechanisms and other advantages, so it is a hot field in genetic breeding research. Regarding the research on the development of turbot SNP markers, before the present invention was made, there were some relevant reports at home and abroad, and the poplar candidate based on EST sequence published by Zhang Xinye, etc. SNPs marker analysis has reported a meth...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 马爱军刘庆明王新安黄智慧
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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