IBD-Appendix
DNA was extracted from 50 mg of appendix tissue collected from 6 IBD patients and 2 non-IBD patients (one with FAP and one with AA) for 16S rRNA sequencing to characterize the bacterial population in the appendix. The extraction was performed using the QIAamp Fast DNA Stool Mini Kit (QIAGEN). The extracted DNA was quantified using a NanoDrop 1000 Spectrophotometer (Thermo Fisher Scientific), and concentrations ranging from 7 to 200 ng/L were used for library preparation. Due to the low detection of 16S amplification under standard conditions (5 ng/L DNA) in the Illumina protocol, higher DNA concentrations (7200 ng/L) were utilized.   Libraries were prepared using the 16S Metagenomic Sequencing Library Preparation protocol (Illumina). The V3V4 region of 16 S rRNA genes was amplified with the KAPA HiFi HotStart ReadyMix kit (Roche Sequencing Solutions) with the following primers: Forward Primer: 5' TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG . Reverse Primer: GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC. Quantification of the libraries was performed using a Qubit 4 Fluorometer (Thermo Fisher Scientific), and their sizes were assessed with the QIAxcel Electrophoresis System (QIAGEN). Each library was diluted to 4 nM, and 5 L of each was pooled for sequencing. The pooled libraries were sequenced on an Illumina MiSeq platform, employing a 2x300 bp paired-end approach, which generated 22-25 million reads in each direction.