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《Molecular Plant Breeding》 2018-10
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Optimization Techniques for SRAP-PCR Reaction System in Mallotus oblongifolius(Miq.) Muell. Arg.

Yan Wuping;Li Juanling;Wu Shumin;Qin Shaochang;Yang Xiangfei;Wang Yu;Institute of Tropical Agriculture and Forestry, Hainan University;  
In order to better use the SRAP molecular marker technology in the study on the genetic diversity of Mallotus oblongifolius, single factor and orthogonal experiments were combined to optimize the main affecting factors in the SRAP-PCR reaction system of Mallotus oblongifolius. First, the single factor experiment was used to find the suitable concentration range of each factor, and then the orthogonal test was carried out by L_(16)(4~4) orthogonal test. According to the requirements of genetic diversity analysis, we directly analyzed the amplification effect between 16 experimental combinations, and constructed a set of SRAP-PCR system with good repeatability, clear bands and stability. The optimum SRAP-PCR reaction system(20 μL) included 2 μL 10×PCR buffer, 3 U Taq DNA polymerase, 0.15 mmol/L d NTPs, 0.4 μmol/L primer and 40 ng DNA template. After verification, this optimum system was not only suitable for the application in the PCR reaction conditions and procedures of SRAP analysis in Mallotus oblongifolius germplasm resources, but also suitable for the analysis of genetic relationship and genetic diversity among different populations in Mallotus oblongifolius.
【Fund】: 国家自然科学基金项目(31560108)资助
【CateGory Index】: S571.1
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