Deep Shotgun Metagenomic Dataset
We weighed 0.2-g portions of 16 freshly thawed fecal samples into 2-ml
bead beating tubes containing Qiagen 0.7-mm garnet beads, added 750 µl
of PowerBead Solution and 60 µl of C1 buffer from Qiagen’s QIAamp
PowerFecal DNA Kits to each tube, vortexed each tube for 2 seconds, and
incubated tubes for 10 min at 60ºC. After incubation, we bead beat
samples for 10 min on a Vortex-Genie 2 fitted with Qiagen’s Vortex
Adapter (cat. no 13000-V1-24). All other extraction steps were completed
as per Qiagen kit recommendations (single-tube extraction protocol).
Final DNA extracts were eluted in 100 µl of molecular grade water.
DNA extracts were quantified using Qubit dsDNA Broad Range Assays.
Extracts were sent to the University of Calgary Centre for Health
Genomics and Informatics, where shotgun metagenomic libraries were
created using v1 Illumina Nextera XT Index Kit preparations, as per
manufacturer specifications. Normalized libraries were sequenced on an
Illumina NextSeq 500 High Output Flow Cell to a target depth of 30
million 2 x 150 bp reads per sample.