| 活性污泥总DNA提取方法的比较 |
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| 引用本文： | 丁嫚,李璐,邹莉,温东辉,唐孝炎.活性污泥总DNA提取方法的比较[J].环境工程学报,2009,3(9):1697-1702. |
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| 作者姓名： | 丁嫚 李璐 邹莉 温东辉 唐孝炎 |
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| 作者单位： | 北京大学环境科学与工程学院,北京,100871 |
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| 基金项目： | 国家自然科学基金资助项目（50878001） |
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| 摘 要： | 从处理某制药废水的MBR反应器中采集活性污泥，评价不同DNA提取方法对其总DNA提取效率的影响。DNA提取分细胞裂解和DNA纯化2步，对细胞裂解比较了珠磨匀浆法、反复冻融法、十二烷基磺酸钠（sodium dodecyl sulfate，SDS）裂解法等7种方法；对DNA纯化比较了酚/氯仿纯化法和胶回收纯化法。结果表明,SDS裂解法、酚氯仿纯化法最优。通过条件优化实验，确定SDS裂解酚/氯仿纯化法在污泥量1.1 g，10 000 r/min离心5 min的操作条件下，获得的DNA产量（10 774 μg/g泥重）和纯度（OD260∶OD280=1.84）等综合指标最好。
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| 关 键 词： | 细胞裂解 DNA纯化 聚合酶链反应(PCR) 变性梯度凝胶电泳(DGGE) 活性污泥 |
Comparison of different DNA extraction methods for activated sludge |
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| Ding Man,Li Lu,Zou Li,Wen Donghui and Tang Xiaoyan.Comparison of different DNA extraction methods for activated sludge[J].Techniques and Equipment for Environmental Pollution Control,2009,3(9):1697-1702. |
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| Authors: | Ding Man Li Lu Zou Li Wen Donghui and Tang Xiaoyan |
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| Institution: | College of Environmental Sciences and Engineering, Peking University, Beijing 100871, China,College of Environmental Sciences and Engineering, Peking University, Beijing 100871, China,College of Environmental Sciences and Engineering, Peking University, Beijing 100871, China,College of Environmental Sciences and Engineering, Peking University, Beijing 100871, China and College of Environmental Sciences and Engineering, Peking University, Beijing 100871, China |
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| Abstract: | Using activated sludge sample,which was collected from an experimental MBR treating pharmacy wastewater,several DNA extraction methods were evaluated.DNA extraction includes two steps of cell lysis and DNA purification.Firstly,seven cell lysis methods,including bead mill homogenization,freeze-thawing,SDS-based method,etc.Sencondly,two purifircation methods,phenol/chloroform purification and agarose gel purification were compared.The results showed that SDS-based method and phenol/chloroform purification wer... |
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| Keywords: | cell lysis DNA purification polymerase chain reaction (PCR) denatured gradient gel electrophoresis (DGGE) activated sludge |
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