454-pyrosequencing

To characterise BCC along the lower WFS, pyrosequencing of the above PCR products was performed at the IBIS/Université Laval Plate-forme d’Analyses Génomiques (Québec, Canada) using the Roche 454 GS-FLX Titanium chemistry. Data analysis was conducted with mothur v.1.39.5 [23]. Raw sequence data was quality trimmed and checked for chimeras, the singletons were removed. Final high-quality sequences were assigned to operational taxonomic units (OTUs) at 97% identity and rarefaction curves were constructed to determine whether sampling depth was sufficient to accurately characterize the BCC. Taxonomic classification was based upon the Greengenes reference database [24] with a 97% bootstrap confidence threshold.

DNA sequences were submitted to the GenBank database as BioProject PRJNA275052.