Metagenomics sputum cystic fibrosis
The library preparation for metagenomics analysis was performed using the kit Nextera DNA Library Prep (Illumina) to randomly fragment the metagenomic DNA by means of transposases activity. The adapters were inserted using the kit Nextera XT Index (Illumina) following the manufacturers instructions. The libraries were quality-assured by removal of residual adapters, exceeding dNTPs and fragments less than 100 bp using magnetic beads AMPure XP (Beckman Coulter). The libraries were quantified by RT-PCR using the kit KAPA-KK4824 Library Quantification Kit (Illumina) following fabricant standards. A final concentration of 4 nM was used to normalize the libraries size to pool them into the sequencing flowcell. The HTS was conducted on NextSeq 500 platform (Illumina) using the running kit NextSeq 500 MID Output (300 cycles) to generate 2x150 bp Paired-End (PE) reads.