Full-length Sequencing of 16S rRNA Gene and Its Analysis Based on the SMRT Sequencing Technology

doi:10.13560/j.cnki.biotech.bull.1985.2017-0036

Abstract

Abstract: Single-molecule sequencing，called as third-generation sequencing technology，is a high-throughput technique developed in last few years. Of them，single-molecule real-time（SMRT）sequencing technology，developed by Pacific BioSciences（PacBio），is the first commercial technology. SMRT sequencing technology could successfully overcome the disadvantage of low accuracy in the third-generation sequencing technology，by generating circular consensus sequence（CCS）through cycle sequencing the template sequence. Therefore，SMRT sequencing technology will allow scientists to profoundly and accurately study the structures and functions of microbial communities in complex environment. Here，we introduced the advantages and disadvantages of SMRT sequencing technology in 16S rRNA gene sequence of microorganism，and summarized the important steps，such as quality control，filtering of error tags，clustering analysis，annotation analysis，etc. of full-length 16S rRNA gene sequence acquired by SMRT sequencing technology. In addition，we pointed out the problems and the feasible solutions while applying SMRT sequencing technology in the study of microbial in complex environment，aiming at providing references for researchers in this field.

Key words: single-molecule real-time sequencing technology, PacBio RS II, third-generation sequencing technology, circular consensus sequence

TANG Yong , LIU Xu. Full-length Sequencing of 16S rRNA Gene and Its Analysis Based on the SMRT Sequencing Technology[J]. Biotechnology Bulletin, 2017, 33(8): 34-39.

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