47-SEY16S
Sequencing libraries of bacterial communities of black band mat of coral reefs, Samples were prepared according to the Illumina 16S Metagenomic Sequencing Library Preparation guide. Briefly, we amplify the variable region 4 of the 16S rRNA gene with the following primers: 515F [For:5'TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGTGCCAGCMGCCGCGGTAA '3] and 806RB [Rev:5'GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGGACTACNVGGGTWTCTAAT '3]. PCRs were performed for all samples with 0.2 M of each primer in a total reaction volume of 25 L using the Qiagen Multiplex PCR Kit. The following cycling conditions were used: 95 C for 15 min, followed by 27 cycles of 94 C for 45 s, 50 C for 60 s, 72 C for 90 s, and a final extension step of 72 C for 10 min. Triplicate samples were pooled and cleaned with ExoProStar 1-Step (GE Healthcare, UK). 16S rRNA gene amplicon libraries were sequenced on the Illumina MiSeq platform (Illumina Inc., San Diego, CA, USA).