Exon conserved sequence amplified polymophic molecular marker and its analysis method

A technology of conserved sequences and molecular markers, applied in the fields of genomics and bioinformatics, can solve the problems of being easily affected by experimental conditions, long process time, unable to provide complete information, etc., to improve repeatability and reliability, reduce time and The effect of low cost and primer design cost

Inactive Publication Date: 2012-05-09
GUANGXI UNIV
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Problems solved by technology

However, with the deepening of research, people found that this technology has certain defects: RAPD is a dominant marker and cannot provide complete information; it is easily affected by experimental conditions; the stability is poor; the results cannot distinguish homozygous and heterozygous
But there are also obvious disadvantages: cumbersome technology, long process time, not allelic fragments detected, expe

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  • Exon conserved sequence amplified polymophic molecular marker and its analysis method
  • Exon conserved sequence amplified polymophic molecular marker and its analysis method
  • Exon conserved sequence amplified polymophic molecular marker and its analysis method

Examples

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Embodiment 1

[0048] In this example, sugarcane variety resources were used as materials, and Arabidopsis thaliana and rice were used as close relatives to carry out molecular markers of exon conserved sequence amplification polymorphisms.

[0049] 1. Select 37 sugarcane variety resources as materials, take heart leaves from sugarcane plants, remove the midrib and cut the middle part for DNA extraction, and use the CTAB method to extract DNA.

[0050] 2. Download the mRNA sequences of Arabidopsis thaliana, sugarcane and rice from the NCBI website, and construct the mRNA databases of these crops respectively. The Arabidopsis mRNA database contains 78,301 records, and the mRNA sequences include complete sequences and partial sequences. The mRNA databases of sugarcane and rice consisted of 75 and 3990 complete mRNA sequences, respectively. Programming software Delphi 7.0 (Borland Software Corporation, USA) was used to program, and four bases (A, T, C, G) were combined to generate 1024 5-base o...

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Abstract

The invention discloses an exon conserved sequence amplified polymorphic molecular marker and its analysis method. According to the invention, mRNA sequence information of known species in public database can be utilized to find out five bases oligonucleotide conserved sequence of gene exon. The conserved sequence is taken as a core to link six bases random sequence to its 3' terminal to form an eleven bases sequence which makes up a primer of 20bp together with a fixed sequence of 5' terminal. Primers with higher occurrence frequency of the eleven bases sequence are preliminarily screened out by mRNA database. Then PCR is carried out for repeatability screening and the FCR based exon conserved sequence amplified polymorphic marker is obtained. The method provided by the invention has advantages of simple operation, high effectiveness, good repeatability, reduced time and cost of primer screening. It is a new effective molecular marking method for the genetic diversity analysis. The method can be widely applied in various fields of genetic map construction, gene or QTL location, marker-assisted breeding, genetic diversity analysis, genome research, mRNA differential expression analysis and the like and has a bright prospect.

Description

technical field [0001] The invention belongs to the fields of genomics and bioinformatics, and relates to a molecular marker of exon conserved sequence amplified polymorphism (Conserved sequence of exon amplified polymorphism, CSEAP) and an analysis method thereof. Background technique [0002] Molecular marker is a genetic marker based on the polymorphism of biological macromolecules (mainly genetic material DNA), which has many advantages such as large number, high polymorphism, not affected by seasons and environments, simple and rapid detection methods, etc. . Molecular markers have become the frontier of life sciences and are widely used in genetic diversity analysis, genetic linkage map construction, germplasm resource identification, gene mapping, kinship analysis, sex identification, genome mapping, gene cloning, library construction and molecular Marker-assisted selection breeding and many other aspects. [0003] Since the concept of molecular markers was first pr...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 何龙飞何海旺许春燕李创珍韦善清
Owner GUANGXI UNIV
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