三疣梭子蟹微卫星多重PCR技术建立及条件的优化

Establishment and optimization of a microsatellite-labeling multiplex PCR system in Portunus trituberculatus

中文关键词: 三疣梭子蟹  SSR  多重PCR体系

英文关键词: Portunus trituberculatus  SSR  Multiplex PCR system

中文摘要:

      选择三疣梭子蟹22个多态性高的微卫星标记，经过引物间的搭配、组合测试以及反应条件的优化（退火温度、Mg2 +含量、dNTPs的浓度等参数），构建了多重PCR扩增技术体系。本研究最终建立了47个二重PCR组合，并在47个二重引物的组合基础上建立15个三重PCR组合，3个四重PCR组合。该多重PCR技术体系的建立为三疣梭子蟹品种选育、种系评估提供快速、有效分子检测技术。

英文摘要:

      By selecting 22 pairs of polymorphic microsatellite markers of Portunus trituberculatus, primers matching, and combinatorial testing, the multiplex PCR amplification system was built and the reaction conditions were optimized, including annealing temperature, Mg2+ concentration, dNTPs concentration and other parameters. Forty-seven combinations of double PCR, 15 combinations of triple PCR, and 3 quadruple PCR combinations were established. The establishment of this technology will provide a fast, effective molecular detecting tool for the P. trituberculatus breeding and germ-line assessment.