三种提取河豚鱼肠道微生物总DNA的方法比较

食品科学 ›› 0, Vol. ›› Issue (): 154-157.

三种提取河豚鱼肠道微生物总DNA的方法比较

李丹，潘迎捷，赵勇，孙晓红*

上海海洋大学食品学院，上海水产品加工及贮藏工程技术研究中心

收稿日期:2011-05-30 修回日期:2012-04-01 出版日期:2012-05-15 发布日期:2012-05-07
通讯作者: 李丹 E-mail:egg19870614@163.com
基金资助:
“创新行动计划”部分地方院校计划项目;上海市教育委员会重点学科建设项目资助

Comparison of Three Methods for Total DNA Extraction of Intestinal Microorganisms in Puffer Fish

Received:2011-05-30 Revised:2012-04-01 Online:2012-05-15 Published:2012-05-07

摘要/Abstract

摘要： 目的：建立一种有效、节约的河豚鱼肠道内容物总DNA提取方法，应用于聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)等分子生物学研究。方法：采取苯酚-氯仿-异戊醇抽提法，QIAamp DNA Stool Mini Kit，改良FastDNA SPIN Kit for Soil这3种DNA提取方法，并结合PCR-DGGE技术分析河豚鱼肠道微生物多样性。结果：苯酚-氯仿-异戊醇抽提法与QIAamp DNA Stool Mini Kit提取的DNA量远高于改良FastDNA SPIN Kit for Soil，并且包含的微生物多样性较高。结论：苯酚-氯仿-异戊醇抽提法作为一种有效、节约的DNA提取方法，可应用于肠道微生物多样性研究。

关键词: 肠道内容物, DNA提取, 微生物多样性, 聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)

Abstract: Objective: To establish an effective and economical method for total DNA extraction from intestinal contents in puffer fish. Methods: Three DNA extraction methods including phenol-chloroform-isoamyl alcohol extraction method, QIAamp DNA Stool Mini Kit, modified FastDNA SPIN Kit for soil were designed and used in combination with polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE) technique to analyze the microbial diversity in the intestine of puffer fish. Results: Much more DNA and higher microbial diversity were extracted by phenol-chloroform-isoamyl alcohol extraction method and QIAamp DNA Stool Mini Kit compared with modified FastDNA SPIN Kit for soil. Conclusion: Phenol-chloroform-isoamyl alcohol extraction method is an effective and economical method for DNA extraction, and can be applied for studying microbial diversity.

Key words: intestinal contents, DNA extraction, microbial diversity, polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE)

中图分类号:

S965.225

李丹，潘迎捷，赵勇，孙晓红*. 三种提取河豚鱼肠道微生物总DNA的方法比较[J]. 食品科学, 0, (): 154-157.

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