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Larimichthys crocea genome-wide SNP and InDel molecular marker method based on double enzyme digestion

A genome-wide, molecular marker technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial assay/inspection, etc., to control the cost of marker development, low cost, and simplify genome building and sequencing.

Active Publication Date: 2016-02-24
JIMEI UNIV
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Problems solved by technology

[0004] The purpose of the present invention is to provide a method that can overcome the problems of uneven distribution and high cost of the existing transcriptome and whole genome resequencing technology in the mining of large yellow croaker molecular markers, and can resequence the entire genome at a lower cost. Carry out high-throughput SNP marker mining based on EcoRI and NlaIII double-digestion SNP and InDel molecular marker method for the whole genome of large yellow croaker to solve the problems raised in the above background technology

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  • Larimichthys crocea genome-wide SNP and InDel molecular marker method based on double enzyme digestion
  • Larimichthys crocea genome-wide SNP and InDel molecular marker method based on double enzyme digestion
  • Larimichthys crocea genome-wide SNP and InDel molecular marker method based on double enzyme digestion

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Embodiment 1

[0029] see Figure 1-3 , a method for whole-genome SNP and InDel molecular markers of large yellow croaker based on double enzyme digestion, comprising the following steps:

[0030] 1) Select experimental materials

[0031] In this experiment, a random group of 96 large yellow croakers at the age of 1 was taken in the same period, with similar body weight and body length ranges; after recording the basic traits of all large yellow croakers, the fin rays were kept, preserved with 70% alcohol by volume, and placed in Store at -20°C for later use.

[0032] 2) Genomic DNA extraction and preservation of large yellow croaker

[0033] a. Grind the frozen fish fins into powder with a mortar in liquid nitrogen, take a centrifuge tube with a specification of 1.5ml, add 1ml of digestion buffer, slowly add the powder into 1ml of digestion buffer, and make the powder in the digestion buffer Wet the surface of the solution evenly, then shake to submerge the sample, and digest at 37°C for...

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Abstract

The invention discloses a larimichthys crocea genome-wide SNP and InDel molecular marker method based on double enzyme digestion. The method comprises the following steps: 1) joint sequence design; 2) genome DNA enzyme digestion; 3) joint sequence connection; 4) sample mixing and PCR amplification; 5) high-throughput sequencing; 6) SNP locus excavation and analysis through sequencing data. According to the method, the modern molecular biology and the advanced high-throughput sequencing technology are combined, the method of combination of EcoRII and NlaIII double enzyme digestion is used, the genome-wide DNA molecules are subjected to enzyme digestion, DNA segments with certain lengths are obtained for carrying out library establishment sequencing and SNP molecular marker excavation, and the uniformly distributed genome-wide SNP locus information is obtained; the workload and cost of genome-wide marker analysis are greatly reduced, and the method is also applicable to the genome-wide marker analysis of other species; the SNP marker excavated through the method is applicable to the study fields such as animal and plant species identification, species genetic genealogy analysis, germplasm resource genetic diversity analysis and genetic breeding.

Description

technical field [0001] The invention relates to the technical field of molecular marker development of the whole genome of large yellow croaker, in particular to a method for molecular markers of SNP and InDel of the whole genome of large yellow croaker based on double enzyme digestion. Background technique [0002] Molecular markers refer to DNA fragments or bases that can reflect a certain difference in the genome between organisms or between populations. It directly reflects the differences between genomic DNA. An important tool; mainly including RFLP (RestrictionFragmentLengthPolymorphism), RAPD (RandomAmplifiedPolymorphicDNA), AFLP (AmplifiedFragmentLengthPolymorphism), [0003] SSR (SimpleSequenceRepeat), SNP (SingleNucleotidePolymorphisms) and InDel (smallInsertandDeletion), etc.; at present, with the development of genotyping technology, in addition to the widespread use of SSR, SNP and InDel markers, other types of markers have been used less and less Although SSR ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6888C12Q2600/156
Inventor 肖世俊王志勇陈俊蔚
Owner JIMEI UNIV
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