| 体外培养瘤胃微生物总DNA的提取与纯化 |
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| 引用本文： | 田晓娟,张志红,黄江丽,黄黄,张国华,王东升,丁建南.体外培养瘤胃微生物总DNA的提取与纯化[J].江西科学,2012,30(1):44-46,102. |
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| 作者姓名： | 田晓娟 张志红 黄江丽 黄黄 张国华 王东升 丁建南 |
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| 作者单位： | 江西省科学院生物资源研究所,江西南昌,330029 |
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| 基金项目： | 江西省科技支撑计划项目（2010BNA09100）; 江西省科学院博士基金项目（2010YYB01 2011YYB05） |
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| 摘 要： | 主要目的是寻找一种能最大限度获取所有类型的瘤胃微生物大片段DNA的提取方法,为后期分子生物学技术研究瘤胃微生物奠定基础。采用液氮冻融+溶菌酶+CTAB和SDS相结合的方法处理瘤胃样品并充分破壁,再采用酚/氯仿/异戊醇抽提、异丙醇沉淀获得DNA粗提物,经过试剂盒纯化后得到瘤胃微生物DNA样品。经特异性引物PCR检测,本方法所提取的DNA包含细菌、古细菌、真菌及原虫四类微生物信息。因此,采用该方法提取的DNA可用于后期实时定量PCR或DGGE等研究。
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| 关 键 词： | 瘤胃微生物 总DNA提取 PCR |
Extraction and Purification of Total DNA from Rumen Microbes in Vitro Fermentation |
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| TIAN Xiao-juan,ZHANG Zhi-hong,HUANG Jiang-li,HUANG Huang,ZHANG Guo-hua,WANG Dong-sheng,DING Jian-nan.Extraction and Purification of Total DNA from Rumen Microbes in Vitro Fermentation[J].Jiangxi Science,2012,30(1):44-46,102. |
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| Authors: | TIAN Xiao-juan ZHANG Zhi-hong HUANG Jiang-li HUANG Huang ZHANG Guo-hua WANG Dong-sheng DING Jian-nan |
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| Institution: | (Institute of Biological Resource of Jiangxi Academy of Sciences,Jiangxi Nanchang 330029 PRC) |
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| Abstract: | This study aimed to find an utmost extraction method of total DNA from rumen microbes in vitro fermentation for the post research on the rumen microbial structure and diversity.The method of liquid nitrogen Freeze-thaw,lysozyme＋CTAB and SDS were combined together in order to break microbal cells for releasing DNA as much as possible.Then phenol/chloroform/Isoamyl alcohol extraction and isopropanol precipitation were used to get the total DNA extracts.The purified DNA was obtained by the DNA purification kit,and identified by PCR experiments.Total DNA extracts included all four kinds of microorgism as bacterium,Archaebacteria,fungi and protozoa.Therefor,DNA extracted by this method can be used in the quantitative-PCR,DGGE and other researchs. |
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| Keywords: | Rumen microorganism Extraction of total DNA PCR |
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