Charybdis feriatus whole-genome microsatellite marker, screening method and application

Abstract

The invention relates to a charybdis feriatus whole-genome microsatellite marker, a screening method and application. The screening method mainly comprises the following steps: extracting charybdis feriatus genome DNA (Deoxyribonucleic Acid); carrying out establishment and high-throughput sequencing on a genome library; carrying out quality control, assembling and evaluation on sequencing data; analyzing and screening microsatellite sites; designing primers of the microsatellite sites; carrying out PCR (Polymerase Chain Reaction) amplification on genome DNA of different individuals of the charybdis feriatus by using the primers; detecting PCR amplification products by using native polyacrylamide gel electrophoresis; determining the genotype of each individual according to different migration distances of the PCR amplification products, thus obtaining a polymorphism graph of heritable variation of the charybdis feriatus. The number of microsatellite site-containing sequences screened and obtained by adopting the screening method is up to 24,101,990; the total number of the microsatellite sites is up to 3,779,209. The screening method disclosed by the invention not only has the advantages of simpleness in operation, fastness, accuracy, sensitiveness, time saving, labor saving, less economic investment and the like, but also is capable of increasing the screening efficiency, and can be applied to analysis on heritable variation of the charybdis feriatus and research on population genetic diversity.

Description

technical field [0001] The invention belongs to the field of microsatellite molecular markers of the rust-spotted catfish, and particularly relates to a whole-genome microsatellite marker of the rust-spotted catfish, a screening method and an application thereof. Background technique [0002] Microsatellites are ubiquitous in eukaryotic genomes and are randomly repeated DNA sequences consisting of 1-6 nucleotides. They are currently a relatively mature molecular marker technology. Microsatellites have the advantages of large number, random distribution, high polymorphism, strong repeatability and co-dominant inheritance, etc., and are widely used in population genetic diversity analysis, germplasm resource protection and management, genetic linkage map construction and QTL positioning etc. [0003] The first step in microsatellite analysis is to obtain microsatellite loci with high information content by various experimental methods. There are many traditional experimental...

AI Technical Summary

Problems solved by technology

Microsatellite markers are ideal markers for research in this area, but there are very few microsatellite markers available at present, which limits the development of related genetic studies

Moreover, PCR amplification can only amplify a certain sequence and cannot cover the entire genome; the randomness of picking clones is very high, and the repetition rate is also high

As a result, in the existing screening method for microsatellite markers of rusty spot moth, PCR amplification is used to clone and sequence the amplified products. Not only is the efficiency low, but the number of microsatellite sites obtained is very small and cannot cover the entire genome

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