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A dna molecular labeling method based on single primer amplification reaction

A DNA molecule, single-primer amplification technology, applied in the biological field, to achieve the effect of simple operation, abundant quantity and high stability

Inactive Publication Date: 2017-12-22
广西壮族自治区农业科学院经济作物研究所
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no DNA molecular labeling technology based on a single primer amplification reaction for the amplification of exon regions of genes.

Method used

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  • A dna molecular labeling method based on single primer amplification reaction
  • A dna molecular labeling method based on single primer amplification reaction
  • A dna molecular labeling method based on single primer amplification reaction

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Embodiment Construction

[0045] The schematic diagram of a DNA molecular labeling technique based on a single primer amplification reaction of the present invention is shown in figure 1 . The single primer used in this technology is designed based on the rich GC in the gene exon region of the eukaryotic genome. During PCR amplification, the single primer can simultaneously anchor and bind to the gene exon region in the genome The PCR product of the sequence between the gene exon binding sites in the eukaryotic genome is obtained to detect the polymorphism between the two binding sites in the gene exon region of the single primer. The polymorphism mainly comes from the following two aspects: one is the polymorphism caused by the point mutation at the primer binding site; the other is the length polymorphism caused by the sequence insertion, deletion and intron length difference between the primer binding sites Sex.

[0046] Hereinafter, the present invention will be further described in detail through th...

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Abstract

The invention discloses an SPAR (single primer amplification reaction) based DNA molecular marking method, which belongs to the field of biotechnology. The method comprises the following steps: designing and synthesizing a single primer, extracting a sample genomic DNA as a template, carrying out PCR amplification in a conventional PCR reaction system and a conventional reaction procedure by using the single primer and the sample DNA template, and after an amplified product is subjected to agarose gel electrophoresis separation, detecting the polymorphism between two binding sites in a genomic gene exon area, wherein the sequence length of the single primer is 17 bp, six base filling sequences at a 5' terminal are AT-base-rich restriction enzyme cleavage site sequences, a mid-core area sequence is randomly composed of 8 GC bases, and 3 selective bases are arranged at the last 3' terminal. The primer in the invention is good in universality, and the DNA molecular marking method disclosed by the invention has the advantages of simple operation, high stability, good repeatability, number abundance, high specificity, high polymorphism, reliable results, and the like.

Description

Technical field [0001] The invention belongs to the field of biotechnology and relates to a DNA molecular labeling method based on a single primer amplification reaction. Background technique [0002] DNA molecular markers are genetic markers based on the variation of nucleotide sequences between individuals, which can directly detect the differences between biological individuals at the DNA level, which can directly reflect the genetic nature of organisms at the DNA level, and are genetic variations at the DNA level of individual organisms. Direct reflection. It has the advantages of abundant quantity, large amount of information, high polymorphism, detection is not affected by seasons, environment and ontogeny stage, and detection methods are simple and rapid. It has been widely used in genetic diversity analysis, kinship evaluation, fingerprinting Construction, genetic linkage map construction, QTL or gene mapping and cloning, molecular marker assisted selection breeding and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6876C12Q2600/156
Inventor 熊发前唐荣华刘俊仙韩柱强贺梁琼唐秀梅蒋菁黄志鹏钟瑞春李忠杨柳史卫东陈忠良邹成林何海旺余功明
Owner 广西壮族自治区农业科学院经济作物研究所
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