Peronia verruculata microsatellite marking and screening method

Abstract

The invention discloses a peronia verruculata microsatellite marking and screening method. The method includes the steps that a peronia verruculata transcriptome sequencing library is built for Illumina HiseqTM2000 paired-end sequencing; based on a sequence obtained through sequencing, analysis and screening are conducted on microsatellite sites through MISA; screening is conducted again through SSRHunter1.3, and it is guaranteed that a front lateral wing and a rear lateral wing of the sequence have enough length for primer design; by means of an SSR primer, genome DNA of a peronia verruculata population is subjected to PCR amplification; amplification products are detected through polyacrylamide gel electrophoresis, and genotypes of the sites are determined according to different migration distances of the amplification products during population detection. Seven polymorphic microsatellite marks are obtained so as to obtain a polymorphic map of peronia verruculata genetic variation. The method is simple and quick, results are true and reliable, and the developed SSR marks can be used for phylogenetic study between peronia verruculata population genetics and onchidium shellfishes.

Description

technical field [0001] The invention belongs to the field of microsatellite molecular markers, in particular to a purple verrucous sulfonic acid microsatellite marker and a screening method. Background technique [0002] The genomes of organisms, especially the genomes of higher organisms, contain a large number of repetitive sequences, which can be divided into tandem repetitive sequences and interspersed repetitive sequences according to the distribution of repetitive sequences in the genome. Among them, the tandem repeat sequence is formed by connecting the first positions of related repeat units and arranging them in series. There are two main types of tandem repeats found so far: one is composed of functional genes (such as rRNA and histone genes); the other is composed of non-functional sequences. Tandem repeats can be classified into satellite DNA, microsatellite DNA, minisatellite DNA and the like according to the length of the repeating unit of the repeating sequen...

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Problems solved by technology

[0004] As a transitional species of marine invertebrates extending to land, the clamshell shellfish of the family Sulphuridae is considered to be the best representative for the study of marine shellfish radiating to land. In the report of genetic diversity, the 7 microsatellite marker sites developed by high-throughput sequencing technology in the present invention not only provide basic data for the study of the genetic diversity of purple verrucoids, but also can be used for the study of the genetic diversity of sulphuridae. In the study of phylogenetic relationships

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