| 一种同时提取水稻土壤中细胞外和细胞内形态DNA的方法 |
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| 引用本文： | 张蕾,徐立新,袁潜华.一种同时提取水稻土壤中细胞外和细胞内形态DNA的方法[J].生物技术通报,2012(1):65-69. |
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| 作者姓名： | 张蕾 徐立新 袁潜华 |
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| 作者单位： | 海南大学农学院,海口,570228 |
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| 基金项目： | 转基因专项，海南研究生创新科研课题 |
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| 摘 要： | 采用灭菌的水稻田土壤为基质,分别投加细菌基因组DNA和细菌活体,应用该方法提取细胞内和细胞外DNA。结果表明,DNA提取效率平均在75.4%-82.3%,DNA纯度OD260/280在1.75-1.85之间。用细菌16S rDNA通用引物PCR表明,DNA纯度能满足PCR要求,细胞内DNA与细胞外DNA互不污染。
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| 关 键 词： | 细胞外DNA 细胞内DNA 水稻土壤 PCR |
A Simultaneous Recovery of Extracellular and Intracellular DNA from Paddy Soil |
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| Zhang Lei
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Xu Lixin
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Yuan Qianhua.A Simultaneous Recovery of Extracellular and Intracellular DNA from Paddy Soil[J].Biotechnology Bulletin,2012(1):65-69. |
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| Authors: | Zhang Lei Xu Lixin Yuan Qianhua |
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| Institution: | Zhang Lei Xu Lixin Yuan Qianhua(College of Agriculture Hainan University,Haikou 570228) |
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| Abstract: | This article provides a method of simultaneous recovery of extracellular and intracellular DNA suitable for molecular studies from paddy soil. With sterile paddy soil as a substrate,dosing bacterial genomic DNA and bacteria in vivo,we applied this method to recover the extracellular and intracellular DNA. The results showed that the DNA extraction efficiency of an average is 75.4%-82.3%,and DNA purity OD260/280 is between 1.75-1.85.A 16S rDNA based PCR technique was evaluated using universal bacterial primers to detect the sample.We obtained evidence that the DNA purity can meet the requirements for PCR.Extracellular and intracellular DNA can avoid any contamination with DNA released by each other. |
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| Keywords: | Extracellular DNA Intracellular DNA Paddy soil PCR |
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