Uninflamed intestinal mucosa microbiome. Severe-recurrent, fistulating, Crohn's disease female patient 191, Rutgeerts score i3, 28 years old. Library batch 2.
16s rRNA V3 and V4 region amplification with specific primers as described in Kozich JJ, Westcott SL, Baxter NT, Highlander SK, Schloss PD. Development of a dual-index sequencing strategy and curation pipeline for analyzing amplicon sequence data on the MiSeq Illumina sequencing platform. Appl Environ Microbiol. 2013 Sep;79(17):5112-20. doi: 10.1128/AEM.01043-13. Epub 2013 Jun 21. PMID: 23793624; PMCID: PMC3753973. Briefly, genomic DNA extracted from human patient intestinal mucosa was subject to PCR. Bacterial specific band was manually excised from polyacrylamide gels to exclude human mitochondrial DNA co-amplified PCR products.