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 forSoil,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。
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-chloroformisoamyl alcohol extraction method is an effective and economical method for DNA extraction, and can be applied for studying microbial diversity.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2012年第9期154-157,共4页
Food Science
基金
上海市科委"创新行动计划"部分地方院校计划项目(08390513900)
上海市科委工程中心建设基金项目(11DZ2280300)
上海市教育委员会重点学科建设项目资助(J50704)
