Shotgun sequencing library preparation
SENSITIVITY
Whole genome shotgun sequencing library was performed the Nextera XT kit and the library was prepared according to the Nextera XT DNA Library Preparation Guide (Illumina). Briefly, DNA concentrations were diluted to 0.2 ng/μl and fragmented by incubating at 55 °C for 7 min. Paired-end Nextera XT indexes were added and 12 cycles of amplification process were completed. Samples were purified with AMPure XP beads according to the manufacturer’s instructions. In order to calculate molarity, DNA concentration was quantified using the Qubit dsDNA High Sensitivity Assay and amplicon size was measured by Agilent Bioanalyser 2100. Lastly, 1 mM of libraries were pooled before loading on the Illumina NextSeq platform for 150 bp paired-end sequencing.