Method for sifting macrobrachium nipponensis microsatellite mark with magnetic bead concentration method

Abstract

The invention provides a method for screening microsatellite markers of Macrobrachium nipponensis by a magnetic beads enrichment method, including the following steps: the extraction of the genomic DNA of the Macrobrachium nipponensis; the hybridization of a probe and a target segment; the enrichment of magnetic beads; positive sequencing clone and the amplification of the genomic DNA according to primers which are related to the flanking sequences of the microsatellite. As the invention combines the magnetic beads enrichment and PRC amplification of bacterial liquid to carry out second-time screening, no isotope is needed. The method has the advantages of safety and high efficiency and low cost, short cycle, high reliability and the like. At the same time, in the enrichment process, the magnetic beads are repeatedly washed for a plurality of times, which can clean and remove microsatellite sequences which are not firmly adhered because of less repetition times. Therefore, the obtained microsatellite sequences are longer and are repeated for more times. The microsatellite sequences which are more frequently repeated are considered to have rich diversity at interspecies and intraspecies and can be widely applied to the diversity identification of germ plasm, the building of genetic maps, and QTL. Therefore, the method is one which screens the microsatellite markers of Macrobrachium nipponensis and can be easily popularized.

Description

technical field [0001] The invention belongs to a DNA molecular genetic marker method of Macrobrachium japonicus, in particular to a method for rapidly separating and screening microsatellite markers of Macrobrachium japonicus by using a magnetic bead enrichment method. Background technique [0002] Macrobrachium nipponensis (Macrobrachium nipponensis), commonly known as freshwater shrimp, belongs to the class Crustacea, Decapod order, Longarm shrimp family, and Macrobrachium genus. It has the characteristics of strong adaptability, wide distribution, miscellaneous food habits, fast growth, and high economic benefits of breeding. It is an important cultured species of freshwater shrimp in my country. Most of the current research work on Macrobrachium japonicus focuses on growth characteristics (Liu Jun et al., 2003), karyotype analysis (Yang Wanxi and Zhou Hong, 2000) and seedling cultivation (He Xugang and Gong Shiyuan, 2003). Due to the limited genetic background data of ...

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Problems solved by technology

Although the isotope sensitivity is high, it is difficult to fully promote due to the high radioactivity, which requires high laboratory protection and is harmful to the human body.