G703_50_PCR_NegControl_S50
Polymerase chain reaction was performed on genomic DNA with the standard 515F and 816R primers- FWD: GTGYCAGCMGCCGCGGTAA; REV: GGACTACNVGGGTWTCTAAT, following Earth Microbiome Project protocols12,13. The PCR products were checked on a 1% agarose gel and cleaned with AmPure magnetic beads (Beckman Coulter, USA) following manufacturers instructions. The quality and quantity of cleaned DNA was assessed with a spectrophotometer (Nanodrop).  Amplicons were transported on ice to the Genome Sciences and Bioinformatics Core of the Penn State College of Medicine, where they were indexed with Illumina barcodes, library was prepared, checked for quality, and 16S sequencing was performed with an Illumina MiSeq at 2x300 bp using V3 chemistry.