sequencing of human metagenome
Bacterial DNA was isolated from crushed clinical samples suspended in Schadler liquid using the commercial QIAamp DNA Stool Mini Kit and QIAamp DNA Mini Kit (Qiagen; Hilden, Germany), according to the manufacturer's protocol. Metagenomic analysis of the intestinal and pancreatic microflora was performed using hypervariable fragments of the 16S rRNA gene. Sequencing was performed using Ion 16SMetagenomics Kit (Thermo Fisher Scientific, Waltham, MA, USA) on the Ion Torrent Personal Genome Machine (PGM) platform. Briefly, DNA was subjected to the amplification of 16S rRNA libraries. The concentration of each 16S library was determined by bioanalyzer Agilent 2100 and High Sensitivity DNA Analysis Kit (Agilent Bioanalyzer, Santa Clara, CA, USA).