metagenome9
The Childrens Hospital of Pennsylvania (CHOP) Microbiome Center conducted DNA sequencing studies to identify bacterial and metagenomic differences between UC and FAP pouch stool. DNA was extracted using the Qiagen DNEasy PowerSoil kit (Germantown, MD).  For 16S rRNA marker gene sequencing, barcoded PCR primers targeting the V1-V2 region of the 16S rRNA gene were used (forward: 5'-AGAGTTTGATCCTGGCTCAG-3', reverse: 5'-TGCTGCCTCCCGTAGGAGT-3').  PCR reactions were carried out in quadruplicate and pooled for sequencing on the Illumina MiSeq instrument, yielding 250bp paired-end sequence reads.