生物技术通报2016，Vol.32Issue(11)：137-143,7.DOI:10.13560/j.cnki.biotech.bull.1985.2016.11.016
宏基因组靶向测序分析皮肤表面微生物群落方法优化
Optimization of Method for Analyzing Microbial Community on Human Skin by Targeted Sequencing of Metagenomics
摘要
Abstract
Based on the experimental design and data analysis, we screened a high-throughput sequencing method which may measure the community of microorganisms in a low level on the human skin as real as possible. By completely random design of the experiments with multi-factor multi-level, we analyzed the effects of varied factors in the DNA extraction, PCR amplification, sample collection and sequencing on the composition characteristics of microbial community. The results showed that regarding the sample collection, the optimal collection swab and extraction liquid were screened out. In the aspect of DNA, the optimal pretreatment method and reagent box type were selected; on PCR reaction, the optimal template concentration and annealing temperature were determined; and on the sequencing aspect, the suitable sequencing depth was also chosen. In conclusion, through above series of experiments to screen the conditions, the appropriate method for analyzing the composition characteristics of community structure of bacteria in a low level was determined, and successfully applied to human palm skin.关键词
皮肤表面微生物群落/宏基因组靶向测序/16S rRNA 基因Key words
microbial community on human skin/targeted sequencing of metagenomics/16S rRNA gene引用本文复制引用
姚雪,刘文丽,裴广倩,童贻刚,罗亚平..宏基因组靶向测序分析皮肤表面微生物群落方法优化[J].生物技术通报,2016,32(11):137-143,7.基金项目
北京市科技新星计划交叉学科合作课题（xxhz201510），上海市刑事科学技术研究院现场物证重点实验室开放课题 （）
