| 简易-高盐沉淀法提取云南松针叶DNA的条件优化 |
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| 引用本文： | 袁晓慧,段安安,许玉兰,刘惠民. 简易-高盐沉淀法提取云南松针叶DNA的条件优化[J]. 安徽农业科学, 2014, 0(15): 4564-4565,4568 |
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| 作者姓名： | 袁晓慧 段安安 许玉兰 刘惠民 |
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| 作者单位： | 西南林业大学林学院,云南昆明,650224;西南林业大学林学院,云南昆明650224;北京林业大学林木育种国家工程实验室林木花卉遗传育种教育部重点实验室,北京100083 |
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| 基金项目： | 西南林业大学科技创新基金(1304) |
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| 摘 要： | [目的]优化简易-高盐沉淀法提取云南松针叶DNA的条件。[方法]采用正交试验设计,对简易-高盐沉淀法提取云南松针叶基因组DNA的提取条件进行优化,然后用浓度1%琼脂糖凝胶电泳检测DNA质量,并用微量紫外分光光度计检测DNA溶液的纯度和浓度,以定性和定量检测为指标,得出简易-高盐沉淀法提取云南松针叶基因组DNA的最佳处理组合。[结果]最佳处理组合为：水浴温度为70℃,V氯仿∶V异戊醇=25∶1,离心时间为8 min,NaCl浓度为2.2 mol/L。[结论]该结果可为开展云南松分子生物学的研究奠定基础。
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| 关 键 词： | 云南松(Pinus yunnanensis) 正交试验 简易-高盐沉淀法 |
Optimization of Extracting DNA from Pinus yunnanensis Needles by Simple-High Salt Precipitation |
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| Affiliation: | YUAN Xiao-hui, LIU Hui-min et al ( Forestry School, Southwest Forestry University, Kunming, Yunnan 650224) |
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| Abstract: | [Objective] To optimize conditions for extracting DNA from Pinus yunnanensis needles by simple-high salt precipitation. [Method] Using orthogonal test,the extraction conditions were optimized. Then,1% of agarose gel electrophoresis was used to detect the quality of the DNA and micro-ultraviolet spectrophotometer was used to detect the purity and concentration of the DNA. The best processing combination that extracting DNA from P. yunnanensis needles by simple-high salt precipitation was decided by qualitative and quantitative indexes. [Result] It is concluded that the optimal combination were as follows： Vchloroform∶Visoamylol= 25∶1,NaCl concentration is 2. 2 mol / L,pyrolysis temperature is 70 ℃ and the centrifuge time is 8 min. [Conclusion]The results will lay a foundation for further research of P. yunnanensis in molecular biology. |
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| Keywords: | Pinus yunnanensis Orthogonal design Simple-high salt precipitation |
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