| 不同提取缓冲液对红树林土壤DNA提取品质的影响 |
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| 引用本文： | 蒋云霞,郑天凌.不同提取缓冲液对红树林土壤DNA提取品质的影响[J].热带海洋学报,2011,30(2). |
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| 作者姓名： | 蒋云霞 郑天凌 |
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| 作者单位： | 1. 南方医科大学公共卫生与热带医学学院环境卫生学系,广东广州,510515
2. 厦门大学生命科学学院,滨海湿地生态系统教育部重点实验室,福建厦门361005;厦门大学近海海洋环境科学国家重点实验室,福建厦门361005 |
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| 基金项目： | 国家高技术研究发展计划项目，国家自然科学基金项目，深港创新圈项目，南方医科大学公共卫生与热带医学学院院长基金项目 |
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| 摘 要： | 针对1种典型的酸性、高有机质含量类型土壤-红树林土壤,从DNA产量、DNA纯度、土壤腐殖酸类物质的提取量以及DNA提取物PCR(polymerase chain reaction)扩增反应的敏感度等方面比较了不同提取缓冲液对土壤DNA品质的影响.结果表明,酸性SDS(sodium dodecyl sulfate)提取缓冲液所获得的土壤DNA产量及纯度均较低;PVP (polyvinylpyrrolidone)提取缓冲液所获得的土壤DNA产量较高,但土壤腐殖酸类物质的提取量亦远高于其他提取缓冲液;酸性SDS提取缓冲液及PVP提取缓冲液所获得的土壤DNA提取物即使稀释到10-3量级亦不能获得目的基因PCR扩增条带.PVPP(polyvinylpolypyrrolidone)提取缓冲液及CTAB(cetyltrimethylammoniumbromide)+PVPP提取缓冲液所获得的土壤DNA产量较PVP提取缓冲液低,但其纯度较高,并且其纯度随提取缓冲液中PVPP浓度的升高而提高;当2%PVPP提取缓冲液及CTAB+PVPP提取缓冲液所获得的土壤DNA提取物分别稀释到10-2及10-1量级时,均能获得PCR扩增条带.综上所述,酸性提取缓冲液及PVP提取缓冲液不适用于酸性、高有机质含量类型土壤DNA的提取,而2%PVPP提取缓冲液及CTAB+PVPP提取缓冲液均能提高此类土壤DNA的提取品质.
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| 关 键 词： | 红树林土壤 DNA提取 DNA产量 腐殖酸 提取缓冲液 |
Evaluation of different extraction buffers on the quality of mangrove soil DNA |
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| JIANG Yun-xia,ZHENG Tian-ling.Evaluation of different extraction buffers on the quality of mangrove soil DNA[J].Journal of Tropical Oceanography,2011,30(2). |
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| Authors: | JIANG Yun-xia ZHENG Tian-ling |
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| Institution: | JIANG Yun-xia 1,ZHENG Tian-ling 2,3 1.Department of Environmental Health,School of Public Health and Tropical Medicine,Southern Medical University,Guangzhou 510515,China,2.Ministry of Education of Key Laboratory for Coast and Wetland Ecosystem,School of Life Sciences,Xiamen Uni-versity,Xiamen 361005,3.State Key Laboratory for Marine Environmental Science,Xiamen University |
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| Abstract: | The authors compared the influence of six different extraction buffers on DNA yields,DNA purity,co-extracted humic compound yields,and their PCR(polymerase chain reaction) sensitivity from a typical acidic and high organic content soil type,mangrove soil.Results showed that the quality of the soil DNA obtained using the acidic SDS(sodium dodecyl sul-fate) buffer was poor.The DNA yields obtained using the PVP(polyvinylpyrrolidone) buffer were higher than the other buff-ers,whereas the co-extracted humic comp... |
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| Keywords: | microbial diversity DNA extraction DNA yield humic compound extraction buffer |
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