摘要： 对影响SRAP反应的4个因素(Ta酶、dNTP、Mg²⁺、引物)4个水平进行正交组合,以建立日本沼虾的SRAP分子标记技术。试验分两步进行:第一步筛选出可有效扩增的引物组合;第二步对筛选出的体系进行优化。结果表明,日本沼虾SRAP反应体系适宜引物组合为Me4Em2,最适条件为在25μL的反应体系中,Mg²⁺、dNTP、Taq酶、引物浓度分别为2.5 mmol/L、0.25 mmol/L、0.64 U/20μL、0.6μmol/L。本研究结果为SRAP分子标记技术在日本沼虾中的应用奠定了基础。

关键词: 日本沼虾, 分子标记, SRAP, 正交设计

Abstract: Four factors ( Taq DNA polymerase,dNTP,Mg²⁺, primer) with four levels were studied by orthogonal design in order to establish SRAP technique of Macrobrachium nipponense. Firstly, effective amplification systems were screened out, which were optimized at the second step. Opt imal amplif icat ion system for Macrobrachium nipponense wasestablished with 2. 5 mmol/ L Taq DNA polymerase, 0.??25 mmol/ L dNTP, 0.64 U/ 20L Mg2+ and 0.6 mol/ L primer.The result laid on foundation for the application of SRAP marker in Macrobrachium nipponense.

Key words: Macrobrachium nipponense, Molecular marker, SRAP, Orthogonal design

中图分类号: 

• S945.1