| 山茱萸叶片DNA提取方法及ISSR反应体系构建 |
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| 引用本文： | 董博文,管少花,董卉卉,赵志浩,李继东.山茱萸叶片DNA提取方法及ISSR反应体系构建[J].河南林业科技,2014(1):1-4. |
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| 作者姓名： | 董博文 管少花 董卉卉 赵志浩 李继东 |
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| 作者单位： | [1]河南农业大学,郑州450002 [2]信阳市林业科学研究所,河南464031 |
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| 基金项目： | 国家林业公益性行业科研专项项目（201104047） |
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| 摘 要： | 山茱萸成熟叶片中多糖及多酚类物质含量较高,传统CTAB法提取其叶片DNA效果较差,该研究在传统的CTAB提取法的基础上进行了改良,经DNA原液电泳检测及PCR产物的电泳检测表明,该法适用于提取硅胶快速干燥保存的山茱萸叶片的DNA。在此基础上,对山茱萸ISSR反应体系中的退火温度、引物浓度及模板用量进行了筛选,结果表明,在25μL的反应体系中,引物UBC847的最佳退火温度为55.2℃,引物最佳浓度为0.8μmol/L,模板最佳用量为70 ng,初步构建了山茱萸ISSR反应体系。
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| 关 键 词： | 山茱萸 DNA提取 ISSR |
A Method for Extracting DNA from Mature Leaves of Comus officinalis and ISSR Reacting System Construction |
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| DONG Bo-wen,GUAN Shao-hua,DONG Hui-hui,ZHAO Zhi-hao,LI Ji-dong.A Method for Extracting DNA from Mature Leaves of Comus officinalis and ISSR Reacting System Construction[J].Journal of Henan Forestry Science and Technology,2014(1):1-4. |
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| Authors: | DONG Bo-wen GUAN Shao-hua DONG Hui-hui ZHAO Zhi-hao LI Ji-dong |
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| Institution: | 1.Henan Agricultural University, Zhengzhou 450002, China; 2. Xinyang Forestry Science Research Institute, Henri 464031, China) |
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| Abstract: | Comus oiticinslis is a kind of medicinal plants with polysaccharide and polyphenol . Conventional eTAB method is useless for extracting DNA of Comus oflicirlCljis.This study improved extraction method based on the Conventional CTAB method, and the result of the spectrophotometry. agarose gel electrophoresis and PCR indicated the improved CTAB method is suitable for the extracting DNA of Comus oiticinslis leaf stored in silica gel.Based on this, the annealing temperature.the concentration of primer and the dosage of DNA template is screened. All our results suggest that the 0.8 μL mol/L primer, 70ng DNA template and 10μL Taq PCR Master Mix in a total volume of 20μL were suitable for ISSR-PCH amplification system of Comus officinalis. The optimal annealing temperature for primer UBC847 is 55.2℃ . |
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| Keywords: | Comus officinalis DNA extraction ISSR |
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