Metagenome sequencing for an anammox bacterium, Candidatus 'Brocadia sinica'. Candidatus Brocadia sinica JPN1

This project was performed to retrieve the genome sequence of an anammox bacterium Candidatus 'Brocadia sinica'. Ca. 'B. sinica' is affiliated into the domain bacteria, the phlum Planctomycetes, order Brocadiales and the genus Candiatus 'Brocadia'. Importantly, all the anammox bacteria were classified into the order Brocadiales while no pure isolate is available (5 June 2012). In the present study, the authors attempted to determine the whole genome sequence of the anammox bacterium, Ca 'B. sinica' by metagenomic approach. Total genomic DNA was extracted from the highly enriched biomass of Ca. 'B. sinica' (>94% in total biomass) and subjected to 454 pyrosequencing with GS FLX titanium. The 454 pyrosequencing was performed with the fragment and 3kb-pair-end sequence techniques and the sequence reads were assembled with GS de novo assembler. The gaps remaining on the scaffolds were closed with the standard primer walking and sanger sequencing techniques.