Exon conserved sequence amplified polymophic molecular marker and its analysis method

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...

AI Technical Summary

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, expensive, and the AFLP marker is a dominant marker, which cannot provide complete information

However, most of them belong to random DNA molecular markers in the traditional sense. The amplified fragments may be non-coding regions, or may be randomly amplified in the genome. Linking to target traits, which makes the application of molecular markers somewhat biased from their targets, thus limiting their application

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