Metagenomics obese vs lean in-vitro dietary fiber
A total of 1 ng of DNA was used following the manufacturers instructions of Nextera XT Library kit (Illumina). DNA was simultaneously fragmented and tagged with dual index sequencing adapters that enable accurate assignment of reads per sample. Library quality control was ensured by profiling and length distribution analysis using the HSD5000kit in the TapeStation 4200 equipment (Agilent). NovaSeq 6000 sequencing platform in a 150 pairedend reads configuration generated *.bcl files as primary sequencing output( NovaSeq Control Software (NCS) version 1.6). Bcl2fastq 2.20 program was used to translate the sequencing reads from bcl (Base Calling) to FASTQ format. This step also removed sequencing adapters. The Clumpify tool from the BBToolssuite (Bushnell B, 2015) was used to remove optical duplicates. Reads with less Phred quality score than Q20 and less length than 50 nucleotides were filtered out using the program BBMap v38.36 (Bushnell B, 2015) Human genome presence was filtered using NGLess v1.0.0Linux64 (Coelho et al., 2019). Homo Sapiens hg19 genome, which is builtin in Ngless was used. We then aligned the sequencesto the genomes and those with alignments of more than 45 bases and 97% similarity were discarded. The remaining sequences were called High Quality sequences and were meant to be the final sequences.