| 文冠果DNA提取及RAPD反应体系的优化 |
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| 引用本文： | 高晓欣,张芸香,郭晋平.文冠果DNA提取及RAPD反应体系的优化[J].山西农业大学学报(自然科学版),2011,31(4):315-320. |
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| 作者姓名： | 高晓欣 张芸香 郭晋平 |
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| 作者单位： | 山西农业大学林学院,山西太谷,030801 |
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| 基金项目： | 山西林业厅科技支撑项目 |
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| 摘 要： | 以山西省20个县的文冠果为材料,采用改良的CTAB法提取文冠果基因组DNA,并对文冠果RAPD分析的最佳反应体系进行优化。结果表明,文冠果RAPD分析的最适反应体系为:PCR扩增的总体积为20μL,包括30ng的模板DNA,10×PCR buffer 2μL,2.0mmol.L-1 Mg2+,0.1mmol.L-1dNTP,Taq酶1U,不足的体积用超纯水补足。扩增程序为:94℃预变性120s,94℃变性30s,36.9℃退火45s,72℃延伸90s,45个循环后在72℃延伸300s,结束后在4℃条件下保存。在此最佳反应条件下,RAPD分析具有良好的稳定性和可重复性。
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| 关 键 词： | 文冠果 RAPD 体系优化 |
Extraction of Xanthoceras Sorbifolia Bunge DNA and Optimization of the RAPD Reaction Conditions |
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| GAO Xiao-xin,ZHANG Yun-xiang,GUO Jin-ping.Extraction of Xanthoceras Sorbifolia Bunge DNA and Optimization of the RAPD Reaction Conditions[J].Journal of Shanxi Agricultural University,2011,31(4):315-320. |
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| Authors: | GAO Xiao-xin ZHANG Yun-xiang GUO Jin-ping |
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| Institution: | GAO Xiao-xin,ZHANG Yun-xiang,GUO Jin-ping(College of Forestry,Shanxi Agricultural University,Taigu Shanxi 030801,China) |
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| Abstract: | The RAPD reaction conditions of Xanthoceras sorbifolia Bunge in 20 counties in Shanxi province were optimized with the modified CTAB method for DNA extraction.The results showed that the optimal RAPD analysis reaction system as： The total volume of PCR amplification 20 μL,including 30 ng of template DNA,10 × PCR buffer 2 μL,2.0 mmol·L-1Mg2＋,0.1 mmol·L-1dNTPs,Taq enzyme 1 U.Amplification program is： 94℃ pre-denaturation for 120 s,94℃ denaturation 30 s,36.9℃ annealing 45 s,72℃ extension 90 s.After 45 cycles,72℃ extension for 300 s,the end stored at 4℃.In optimum conditions,RAPD analysis has good stability and repeatability. |
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| Keywords: | Xanthoceras sorbifolia Bunge RAPD System optimization |
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