Illumina MiSeq paired end sequencing of SSUB007569 : 7
The obtained amplicons were extracted from 2% agarose gels and purified using the AxyPrep DNA Gel Extraction Kit (Axygen Bioscience, Union City, CA, U.S.) according to the manufacturer??????s instructions and quantified using QuantiFluorTM ??????ST (Promega, U.S.). Purified amplicons were pooled in equimolar and paired-end sequenced (2 ???? 300) on an Illumina MiSeq platform according to the standard protocols. The extraction of high-quality sequences was firstly performed with the QIIME package (Quantitative Insights Into Microbial Ecology) (v1.2.1). The remaining forward and reverse sequencing reads were then fused with each other by the program PEAR and FLASH. Reads which could not be assembled were discarded. The unique sequence set was classified into operational taxonomic units (OTUs) under the threshold of 97% identity using UCLUST. Chimeric sequences were identified and removed using Usearch (vertion 8.0.1623). The taxonomy of each 16S rRNA gene sequence was analyzed by UCLUST against the Silva119 16S rRNA database using confidence threshold of 90%.