Metagenomic shotgun sequencing of homo sapiens tracheobronchial secretions
Oropharyngeal swabs and stool samples were collected in collection tubes containing DNA/RNA shield (Zymo Research Cat# R1107-E and Cat# R1101) and frozen at -80C until further analysis was performed. DNA was isolated from the oropharyngeal swabs and stool samples using the ZymoBIOMICS DNA Miniprep Kit (Cat#D4300). For DNA isolation lysis of microbes was performed by mechanical disruption using a Mini-BeadBeater-96 (BioSpec) two times for 2 min. Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit (NEB Biolabs) according to manufacturer's instructions. Sequencing was performed on the Illumina NovaSeq platform (PE150).