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SPAR (single primer amplification reaction) based DNA molecular marking method

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

Inactive Publication Date: 2015-09-30
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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  • SPAR (single primer amplification reaction) based DNA molecular marking method
  • SPAR (single primer amplification reaction) based DNA molecular marking method
  • SPAR (single primer amplification reaction) based DNA molecular marking method

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Embodiment Construction

[0045] A schematic diagram of DNA molecular labeling technology based on single primer amplification reaction of the present invention is shown in figure 1 . A single primer used in this technology is designed according to the rich GC in the exon region of the eukaryotic genome. During PCR amplification, the single primer can be anchored and combined in the exon region of the genome at the same time To obtain the PCR product of the sequence between the exon binding sites in the eukaryotic genome, so as to detect the polymorphism between the two binding sites of the single primer in the exon region of the genome. Polymorphism mainly comes from the following two aspects: one is polymorphism caused by point mutation at the primer binding site; the other is length polymorphism caused by sequence insertion, deletion and intron length difference between primer binding sites sex.

[0046] Below by the embodiment of 16 kinds of peanuts, 8 kinds of sugar cane, 11 kinds of corn, 9 kin...

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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 nucleotide sequence variation among individuals. They can directly detect differences between biological individuals at the DNA level, and can directly reflect the genetic nature of organisms at the DNA level. They are genetic variations at the DNA level of individual organisms. a direct reflection of. It has the advantages of rich quantity, large amount of information, high polymorphism, detection is not affected by season, environment and individual developmental stage, simple and rapid detection method, etc., and has been widely used in genetic diversity analysis, kinship evaluation, fingerprint map Construction, genetic linkage map construction, QTL or gene mapping and cloning, and molecular marker-assisted selection bree...

Claims

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Application Information

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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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