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罗汉果SRAP反应体系的建立与优化 |
刘丽华1, 马小军1*, 孙晶晶2, 覃嘉明3, 魏建和1
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1.中国医学科学院 北京协和医学院 药用植物研究所, 北京 100193;2.辽宁工程
技术大学 理学院, 阜新 123000;3.广西玉米研究所, 南宁 530227
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摘要: |
建立适合罗汉果的SRAP-PCR 扩增体系,为罗汉果的遗传图谱构建及基因定位奠定基础。实验对罗汉果SRAP-PCR 反应体系的影响因素(引物,dNTP,Taq酶,Mg2+,模板DNA)在多个水平上进行优化试验,筛选出各反应因素的最佳水平,建立了罗汉果SRAP-PCR反应的最佳体系(10 μL):引物0.6 μmol/L、dNTP 0.25 mmol/L、Taq DNA聚合酶0.5U、Mg2+ 2.0 mmol/L和模板DNA 30 ng。该体系的建立能很好的满足罗汉果基因组DNA的扩增要求,SRAP标记应用于罗汉果遗传研究是可行的。 |
关键词: 罗汉果 SRAP-PCR 反应体系 建立 |
DOI: |
分类号:Q943.2 |
Fund project: |
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Establishment and optimization of SRAP reaction system in Siraitia grosvenorii |
LIU Li-Hua1, MA Xiao-Jun1, SUN Jing-Jing2,
QIN Jia-Ming3, WEI Jian-He1
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1.Institute of Medicinal Plant Development, China Academy Medicinal Science, Chinese Peking Union Medical
Colledge, Beijing 100193, China;2.College of Science, Liaoning Technical University, Fuxin 123000,
China;3.Guangxi Maize Research Institute, Nanning 530227, China
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Abstract: |
The design was used to establish SRAP-PCR amplification system on Siraitia grosvenorii DNA so as to lay foundation for genetic map construction and gene mapping. The factors influencing SRAP analysis,including primers,dNTP,Taq polymerase,Mg2+,and the concentration of DNA template were studied in a number of levels respectively. The results showed that: 0.6 μmol/L primer,0.25 mmol/L dNTP,0.5U Taq DNA polymerase,2.0 mmol/L Mg2+ and 30 ng DNA template in 10 μL reaction system were the best suitable SRAP-PCR system for Siraitia grosvenorii. It was feasible to apply SRAP marker in genetic research in Siraitia grosvenorii. |
Key words: Siraitia grosvenorii SRAP-PCR reaction system establishment |
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