摘要
探究在不借助其它仪器设备和试剂的条件下,土壤总DNA提取的优化方案。结合荧光定量PCR技术和高通量测序技术对土壤总DNA的提取次数进行研究;发现3次提取的DNA样品的核酸累计量和16S rRNA基因的拷贝数分别占总量的94.84%、85.37%,前4次提取的DNA样品的测序分析结果显示各样品中细菌的相对丰度差异不显著。因此,建议绝对定量分析的实验进行2次以上的DNA提取,定性分析和相对定量分析实验则只需1次提取即可。
To explore an optimization soil total DNA extraction method without the help of other instruments or reagents,real-time PCR and high throughput sequencing was applied to research on the DNA extraction times. The results showed that three times extraction could get more than 94. 84% of total nucleic acids and 85. 37% of total 16 S rRNA gene copies. However,quartic successive extractions did not significantly change the relative abundance of mail bacteria after sequencing. So,we suggest more than double extractions being taken when making a quantitative research,conversely,one time is enough.
作者
陆文利
杨超
颜卫东
LU Wen-li, YANG Chao, YAN Wei-dong(College of Horticultural Science and Engineering, Shandong Agricultural University, Taian 271018, China)
出处
《实验室科学》
2018年第3期57-59,63,共4页
Laboratory Science
基金
山东农业大学教研课题(项目编号:X2017124)
关键词
定量PCR
土壤总DNA
DNA提取
fluorogenic quantitative PCR
soil total DNA
DNA extraction
