Bioo nextflex 96 adapter set

Urban sewage was collected globally from 79 sample locations, covering seven geographical regions from 74 cities in 60 countries^{17 (link)}. DNA was extracted from the sewage pellets according to an optimized protocol using the QIAamp Fast DNA Stool Mini Kit including twice the input material and initial bead beating⁵⁷ . DNA from all samples was mechanically sheared to a targeted fragment size of 300 bp using ultrasonication (Covaris E220evolution). Library preparation was performed with the NEXTflex PCR-free Library Preparation Kit (Bioo Scientific). The Bioo NEXTflex-96 adapter set (Bioo Scientific) was used, and in batches of roughly 60 samples, the libraries were multiplexed and sequenced on the HiSeq. 3000 platform (Illumina), using 2 × 150-bp paired-end sequencing per flow cell with a mean of 120 million reads (range: 8 to 398 million) per sample.

DNA was shipped on dry ice for library preparation and sequencing to the Oklahoma Medical Research Foundation (OMRF). DNA from all samples was mechanically sheared to a targeted fragment size of 300 bp using ultrasonication (Covaris E220evolution). Library preparation was performed with the NEXTflex PCR-free Library Preparation Kit (Bioo Scientific). The Bioo NEXTflex-96 adapter set (Bioo Scientific) was used, and in batches of roughly 60 samples, the libraries were multiplexed and sequenced on the HiSeq3000 platform (Illumina), using 2 × 150-bp paired-end sequencing per flow cell. The raw sequencing data have been deposited at the European Nucleotide Archive under accession number ERP015409.