| 4种镰刀菌基因组DNA提取方法的比较研究 |
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| 引用本文： | 张宝俊,李志岗,张家榕,王建明.4种镰刀菌基因组DNA提取方法的比较研究[J].安徽农业科学,2008,36(31). |
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| 作者姓名： | 张宝俊 李志岗 张家榕 王建明 |
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| 作者单位： | 山西农业大学农学院,山西太谷,030801;山西农业大学农学院,山西太谷,030801;山西农业大学农学院,山西太谷,030801;山西农业大学农学院,山西太谷,030801 |
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| 基金项目： | 山西省自然科学基金
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山西省留学回国人员科研启动基金
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| 摘 要： | 目的]寻找一种简便、高效的基因组DNA提取方法,为进一步开展镰刀菌等丝状真菌的分子生物学研究提供科学基础。方法]采用改良的CTABS、DS、高盐沉淀和SDS-CTAB 4种方法提取8个镰刀菌菌株的基因组DNA,并对其进行质量测定及PCR分析,比较不同提取方法的效果。结果]4种提取方法均可提取到基因组DNA,其抽提质量优劣依次为SDS、SDS-CTAB法、高盐沉淀法和CTAB法。其中采用SDS法可成功提取到所有供试菌株基因组DNA,而且DNA浓度和纯度均较高,RNA及其他杂质污染少。而高盐沉淀法和CTAB法提取到的基因组DNA不够完整或浓度较低。结论]SDS法更适合于镰刀菌等丝状真菌基因组DNA的提取。
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| 关 键 词： | 镰刀菌 基因组DNA 提取方法 |
Comparative Study on 4 Kinds of Extraction Methods of Genomic DNA from Fusarium spp |
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| Abstract: | Objective] The aim was to search a convenient and high efficient extraction method for genomic DNA to provide the scientific basis for further developing the molecular biological study of filamentous fungi such as Fusarium spp.Method] The genomic DNA from 8 Fusarium spp.strains were extracted by 4 methods of improved CTAB,SDS,high salt precipitation and SDS-CTAB and the effects of the different extraction methods were compared through the quality detection and PCR analysis.Result] The 4 extraction methods all could extract the genomic DNA and their order of the extraction quality from good to bad was SDS,SDS-CTAB,high salt precipitation and CTAB.Among them,the SDS method could extract the genomic DNA of all the tested strains successfully and the concn.and the purity of the DNA were all higher,the pollution by RNA and other impurity was little.While the genomic DNA extracted by high salt precipitation and CTAB were not integrity or its concn.was lower.Conclusion] The SDS method was more suitable for the extraction of genomic DNA from filamentous fungi such as Fusarium spp.. |
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| Keywords: | Fusarium spp Genomic DNA Extraction method |
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