title: WGS Illumina sequencing of Fusobacterium nucleatum str. 12230
design: A glycerol stock of F. nucleatum strain 12230, provided by the lab of Yiping W. Han, was streaked on a pre-reduced Columbia blood agar plate, which was then incubated anaerobically at 37C. 10mL of pre-reduced Columbia broth was inoculated with a colony from this plate and incubated for 24 hours at 37C. A 3mL aliquot of the 24 hour Columbia broth culture was prepared according to the DNeasy Blood and Tissue Kit for Gram Negative Bacterial (Qiagen) to isolate DNA. The amount of DNA isolated was quantified using the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen). The 24 hour time point was chosen based on previous experiments showing that cells in a 10mL culture were still dividing at 24 hours with an average optical density (OD) of 0.62 for two replicates. The OD of the cultures peaked at 27 hours at an average of 0.82, but then dropped off slightly at 30 hours with an average OD of 0.68. A 24 hours sample was taken to avoid this drop off point.  Sequencing libraries were prepared as described in the Nextera XT DNA Sample Preparation Guide (Illumina). Library normalization and pooling was performed at the Donnelly Sequencing Center at the University of Toronto.