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Position: Home > Articles > Orthogonal Design and Optimization of SRAP Amplification System for Oncorhynchus keta Chinese Journal of Fisheries 2010,23 (2) 10-14

大麻哈鱼SRAP反应体系正交设计及优化

作  者:
张慧;孙中武;刘伟;尹洪滨
单  位:
中国水产科学研究院黑龙江水产研究所;东北林业大学生命科学学院
关键词:
大麻哈鱼;SRAP;正交设计;单因子试验
摘  要:
以大麻哈鱼基因组DNA为模板,利用相关序列多态性(SRAP)技术以及L25(56)正交试验和单因子试验方法,分析了不同浓度的TaqDNA聚合酶、引物、dNTPs、Mg2+、DNA模板对扩增结果的影响,最终确立的PCR最佳反应体系为15μl,该体系含有1×PCRbuffer,TaqDNA聚合酶0.75U,引物浓度0.3μmol/L,dNTPs浓度0.3mmol/L,Mg2+浓度2.5mmol/L,DNA模板100ng。利用此优化反应体系,获得了清晰、稳定、多态性高的DNA谱带。
译  名:
Orthogonal Design and Optimization of SRAP Amplification System for Oncorhynchus keta
作  者:
ZHANG Hui1,2,SUN Zhong-wu1,LIU Wei2,YIN Hong-bin2 (1.College of Life Sciences,Northeast Forestry University,Harbin 150040,China;2.Heilongjiang River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Harbin 150070,China)
关键词:
Oncorhynchus keta;SRAP;Orthogonal design;Single factor test
摘  要:
Effects of different concentrations of Taq DNA polymerase,primer,dNTPs,Mg2+,DNA template on amplification result were studied by single factor test,orthogonal design of L25(56) and Sequence related amplified polymorphism marker(SRAP) technique,using the genomic DNA extracted from Oncorhynchus keta as template.The optimal PCR system was obtained as 1×PCR buffer,0.75U Taq DNA polymerase,0.3μmol/L primer,0.3mmol/L dNTPs,2.5mmol/L Mg2+,and 100ng genomic DNA in 15μl reaction system.Under the optimal reaction system,we have obtained clear,stable and abundant polymorphic bands.

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