Optimization of Method for Analyzing Microbial Community on Human Skin by Targeted Sequencing of Metagenomics

doi:10.13560/j.cnki.biotech.bull.1985.2016.11.016

Abstract

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.

Key words: microbial community on human skin, targeted sequencing of metagenomics, 16S rRNA gene

YAO Xue, LIU Wen-li, PEI Guang-qian, TONG Yi-gang, LUO Ya-ping. Optimization of Method for Analyzing Microbial Community on Human Skin by Targeted Sequencing of Metagenomics[J]. Biotechnology Bulletin, 2016, 32(11): 137-143.

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