Shotgun Metagenome of homo sapiens: adult DDEB skin
DNA was extracted from swabs that were transferred into Lysing Matrix E tubes (MP Biomedicals) with 500 l of ATL Buffer (Qiagen). Samples were subjected to bead-beating in a FastPrep-24 Instrument (MP Biomedicals) at 6.0 m/s for 40 s and then centrifuged at 16,000g for 5 min. 200 L of the supernatant were treated with Proteinase K (Qiagen) and incubated at 56 C for 15 min before extraction in the EZ1 Advanced XL Instrument (Qiagen) using the EZ1 DNA Tissue Kit (Qiagen) according to the manufacturers protocol. DNA concentration was quantified using a Qubit fluorometer and the Qubit dsDNA HS Assay Kit (Life Technologies). DNA libraries were constructed with 50 L of extracted DNA after shearing by Adaptive Focused AcousticsTM technology (Covaris) using the GeneRead DNA Library I Core Kit (Qiagen). Custom index primers were used for the enrichment of DNA libraries with 14 cycles of PCR. Libraries were quantified using the Agilent DNA 1000 Kit (Agilent Technologies) on an Agilent 2100 Bioanalyzer (Agilent Technologies), normalized and pooled in equimolar concentrations for 101 bp paired-end sequencing on an Illumina HiSeq 2000 Instrument at the Next Generation Sequencing Platform (Genome Institute of Singapore). Before uploading the sequencing reads, any reads mapping to the human genome were removed from the dataset. Reads were mapped to the human reference genome (GRCh38) using BWA-MEM (version 0.7.10-r789) with default settings.