metagenomic shotgun sequencing of human feces: PD subject 16
A total amount of 1g DNA per sample was used as input material for the DNA sample preparations. Sequencing libraries were generated using NEBNext Ultra DNA Library Prep Kit for Illumina (NEB, USA) following manufacturers recommendations, The genomic DNA sample was fragmented by sonication to a size of 350 bp, end-polished, A-tailed, ligated with the full-length adaptor for Illumina sequencing, followed by PCR amplification with P5 and indexed P7 oligo adaptor sequences. Finally, PCR products were purified (AMPure XP system) and libraries were analyzed for size distribution by Agilent2100 Bioanalyzer (Agilent Technologies, CA, USA) and quantified using real-time PCR. The Illumina libraries were sequenced on a Illumina NovaSeq 6000 (150 bp paired-end). Reads containing adapters sequences, 10% of undetermined bases and low-quality bases were removed.