title: 16S rRNA library of human microbiome: stool sample
design: We used Ion 16STM Metagenomics Kit (Life Technologies, Carlsbad, CA, USA) to amplify V2-4-8 and V3-6, V7-9 regions of 16S rRNA in two separate PCR reactions. Amplified DNA was purified using Agencourt AMPure XP (Beckman Coulter, Brea, CA, USA). Purified PCR products were quantified on LabChip GX TouchTM using HT DNA High Sensitivity Reagent Kit (both PerkinElmer, Waltham, MA, USA). Libraries were prepared using Ion Plus Fragment Library Kit and barcode adapters Ion Xpress TM Kit (both Life Technologies, Carlsbad, CA, USA). Prior to chip loading, samples were processed using Ion ChefTM instrument and Ion 510 & amp, Ion 520 & amp and Ion 530 Chef Kits (all Life Technologies, Carlsbad, CA, USA). Processed samples were loaded onto Ion 350TM chip and sequenced using Ion S5TM system (Life Technologies, Carlsbad, CA, USA). Each sample was sequenced once.