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油葵SSR反应体系的优化
引用本文:艾鹏飞,赵治国,方闪闪.油葵SSR反应体系的优化[J].河北机电学院学报,2010(1):4-7.
作者姓名:艾鹏飞  赵治国  方闪闪
作者单位:河北科技大学生物科学与工程学院,河北石家庄050018
基金项目:河北科技大学科技创新基金资助项目
摘    要:采用改良CTAB法从油葵种仁中提取基因组DNA,用于SSR—PCR反应。对影响油葵SSR—PCR反应的几个重要因子进行了探讨,确定了最佳的反应体系:在总体积为25uL中,DNA为2.0ng/uL,Mg^2+浓度为1.5mmol/L,dNTPs浓度为0.2mmol/L,Taq酶量为2.0U,引物浓度为0.25umol/L,退火温度为52-54℃,扩增的谱带清晰、多态性丰富。该SSR—PCR反应体系为油葵亲缘关系分析奠定了技术基础。

关 键 词:油葵  DNA  SSR—PCR  优化

Optimization of SSR analysis system in oil sunflower
Authors:AI Peng-fei  ZHAO Zhi-guo  FANG Shan-shan
Affiliation:(College of Bioscience and Engineering, Hebei University of Science and Technology, Shijiazhuang Hebei 050018, China)
Abstract:In this study, the top-quality genomic DNA was extracted from seeds of oil sunflower with an improved method of CTAB, and the SSR analysis system was optimized. The factors, which vitally affected the SSR results were studied. And the optimal results are as follows: DNA 2.0 ng/uL, Mg^2+ 1.5 mmol/L, dNTPs 0.2 mmol/L, Taq polymerase 2.0 U and primers 0.25 umol/L in reaction mixture of 25 uL and annealing temperature 52-54℃. Using the above PCR system, SSR pattern was clear and abundant. Thus, the optimal SSR analysis system provided some technical foundations to study the phylogenetie relationship of oil sunflower.
Keywords:oil sunflower  DNA  SSR-PCR  optimization
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