一种提取鱼肠道微生物DNA的新方法的建立并与其它两种方法比较
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- 英文论文题名：A novel genomic DNA extraction method for fish intestinal microflora and comparation with other methods though 16S rDNA high-throughput sequencing
- 论文作者：韩卓然 ; 孙敬锋
- 英文论文作者：Zhuoran Han ; Jingfeng Sun ; Tianjin key Lab of Aqua-ecology and Aquaculture ; Fisheries College ; Tianjin Agricultural University
- 年：2016
- 作者机构：天津农学院水产学院天津市水产生态及养殖重点实验室;
- 论文关键词：肠道微生物 ; 锦鲤 ; DNA提取 ; 高通量测序
- 英文论文关键词：intestinal microflora ; Cyprinus carpio ; DNA extraction ; high-throughput sequencing
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.1
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：551k
- 原文格式：D
- 会议级别：全国
摘要
本实验建立了一种提取鱼肠道微生物DNA的新方法(CLU),此方法高效,经济且简单。CLU法与氧化锆珠破壁法(ZBC)及Qiagen试剂盒(QIA)两种方法同时提取锦鲤肠道微生物DNA,并通过琼脂糖凝胶电泳及16S rDNA高通量测序进行检测及比较。在方法CLU中,将溶菌酶与超声波结合分离基因组DNA,且首次将溶菌酶的孵育温度提高到60℃。琼脂糖凝胶电泳表明CLU法和QIA法可得到单一清晰的DNA条带;方法 ZBC法得到的DNA虽降解严重,但得率高。16S rDNA高通量测序结果表明,CLU法所提取DNA鉴定出的OUT数目显著高于其他两种方法。α-多样性指数表明CLU法提取的DNA生物多样性最高,QIA法最低。ZBC法与QIA法间的微生物组成结构与CLU和ZBC间,或CLU与QIA间相比,更加相似。
The purpose of this study was to describe an efficient, economic and simple method(combination of lysozymeand utrasonic lysis method, CLU) for extracting genomicDNA from Cyprinus carpio intestinal microbial and compare it with Zirmil-beating cell disruption method(ZBC) and QIAamp Fast DNA Stool Mini Kit(QIA) based on agarose gel electrophoresis and 16 S rDNA high-throughput sequencing. In method CLU, the combination of lysozymeand utrasonic lysis was used to extract genomicDNA and the incubation temperature of 60℃ was used for the step of lysozyme lysis for the first time. Agarose gel electrophoresis showed that genomicDNA extracted using the CLU and QIA method made a clear single band. The DNA band obtained by ZBC was degraded severely, but showed a good DNA yield. The result of 16 S rDNA high-throughput sequencing showed that the numbers of OTUs was higher for method CLU than those for the methods ZBC and QIA. T
The purpose of this study was to describe an efficient, economic and simple method(combination of lysozymeand utrasonic lysis method, CLU) for extracting genomicDNA from Cyprinus carpio intestinal microbial and compare it with Zirmil-beating cell disruption method(ZBC) and QIAamp Fast DNA Stool Mini Kit(QIA) based on agarose gel electrophoresis and 16 S rDNA high-throughput sequencing. In method CLU, the combination of lysozymeand utrasonic lysis was used to extract genomicDNA and the incubation temperature of 60℃ was used for the step of lysozyme lysis for the first time. Agarose gel electrophoresis showed that genomicDNA extracted using the CLU and QIA method made a clear single band. The DNA band obtained by ZBC was degraded severely, but showed a good DNA yield. The result of 16 S rDNA high-throughput sequencing showed that the numbers of OTUs was higher for method CLU than those for the methods ZBC and QIA. T
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