Low microbial diversity and abnormal microbial succession is associated with Necrotizing Enterocolitis in preterm infants
Microbial DNA was isolated from meconium and fecal samples using the QIAamp Fast DNA Stool Mini Kit (Qiagen, Valencia, CA, USA) following the manufacturer's instructions. DNA quality was determined by spectrophotometry using NanoVue™ spectrophotometer (GE Healthcare, Chicago, IL, USA). All DNA samples were stored at -80 °C until use. The V4 region of the 16S rRNA gene was amplified and sequenced using the PGM Ion Torrent (Thermo Fisher Scientific, Waltham, MA, USA) with the bacterial/archaeal primers 515F and 806R. The reactions were combined in equimolar concentrations to create a mixture composed by 16S gene amplified fragments of each sample. This composite sample was used for library preparation with Ion OneTouch™ 2 System with the Ion PGM™ Template OT2 400 Kit Template (Thermo Fisher Scientific, Waltham, MA, USA). The sequencing was performed using Ion PGM™ Sequencing 400 on Ion PGM™ System using Ion 318™ Chip v2 with a maximum of 40 samples per microchip.
Unipampa
Metagenome of  preterm infants with Necrotizing Enterocolitis