16S rRNA sequencing of DSS(-) WT mice_5
A 16S rRNA sequencing library was constructed according to the 16S metagenomics sequencing library preparation protocol (Illumina, San Diego, CA, USA) targeting the V3 and V4 hypervariable regions of the 20S rRNA gene. KAPA HiFi HotStart ReadyMix (KAPA Bistems, Wilmington, MA, USA) and Agencourt AMPure XP system (Beckman Coulter Genomics, Brea, CA, USA) were used for PCR and purification of the PCR product, respectively. Genomic DNA was isolated from intestine of abalone using PowerSoil DNA Isoloation kit (Mo Bio). The initial PCR was performed with 12 ng template using region-specific primers shown to have compatibility with Illumina index and sequencing adapters (forward primer: 5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGTCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG-3; reverse primer: 5-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC-3). After magnetic bead-based purification of PCR products, the second PCR was performed using primers from a Nextera XT Index Kit (Illumina) with a limited cycle.