| 天牛基因组DNA提取方法和标本保存方式的比较研究 |
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| 引用本文： | 谭亮魁,王文凯.天牛基因组DNA提取方法和标本保存方式的比较研究[J].安徽农业科学,2008,36(12):4869-4872. |
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| 作者姓名： | 谭亮魁 王文凯 |
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| 作者单位： | 长江大学农学院,湖北荆州,434025;长江大学农学院,湖北荆州,434025 |
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| 基金项目： | 国家自然科学基金"中国沟胫天牛亚科比较形态学和系统分类学研究"(30570196). |
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| 摘 要： | 目的]为天牛的分子系统学研究奠定基础。方法]采用不同提取方法(饱和NaCl法、CTAB法、SDS-蛋白酶K消化法)从不同保存条件下天牛标本中提取基因组DNA,进行RAPD扩增,比较不同保存条件和不同DNA提取方法对DNA提取质量的影响,探索天牛标本的最佳保存方式和最佳DNA提取方法。结果]不同方法提取的DNA质量有明显差异。CTAB法和SDS-蛋白酶K消化法提取的DNA质量较好,条带清晰完整,无降解和RNA污染。但饱和NaCl法的提取效果较差。天牛标本的最佳保存条件为在-80℃条件下无水乙醇浸泡。CTAB法提取的天牛基因组DNA能扩增出稳定清晰的多态性片段。结论]CTAB法提取的天牛DNA质量最好,可直接用于PCR扩增。
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| 关 键 词： | 天牛 DNA提取 保存方式 |
| 文章编号： | 0517-6611(2008)12-04869-03 |
| 修稿时间： | 2008年3月7日 |
Comparative Study on Different Extracting Methods of Genomic DNA from Cerambycidae and Different Samples Preservation Modes |
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| TAN Liang-kui.Comparative Study on Different Extracting Methods of Genomic DNA from Cerambycidae and Different Samples Preservation Modes[J].Journal of Anhui Agricultural Sciences,2008,36(12):4869-4872. |
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| Authors: | TAN Liang-kui |
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| Abstract: | Objective] The aim of the research was to lay the foundation for the molecular systematics study of Cerambycidae.Method] Genomic DNA was extracted from Cerambycidae samples under different preservation conditions by different extracting methods(saturated NaCl method,CTAB method and SDS-proteinase K Method) to make RAPD amplification.The effects of different preservation conditions and different DNA extracting methods on the extraction quality of DNA were compared.And the optimum preservation mode of Cerambycidae samples and the optimum DNA extracting method were explored.Result] The quality of DNA extracted by different methods had obvious difference.The quality of DNA extracted by CTAB method and SDS-proteinase K Method was better with clear and clean bands,without degradation and RNA population.But the extracting effect by saturated NaCl method was worse.The optimum preservation conditions of Cerambycidae samples were soaking with anhydrous alcohol at-80 ℃.The genomic DNA extracted from Cerambycidae by CTAB method could amplify stable and clear polymorphic fragments.Conclusion] The quality of DNA extracted by CTAB method was best and it could be directly used in PCR amplification. |
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| Keywords: | Longhorn beetles DNA extraction Preservation mode |
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