GSM7775161: Il22ra1f/f-Colon feces, Fc, replicate 9; feces metagenome; OTHER
The UC San Diego Microbiome Core performed nucleic acid extractions from feces utilizing previously published protocols6. Briefly, samples were purified using the MagMAX Microbiome Ultra Nucleic Acid Isolation Kit (Thermo Fisher Scientific, USA) and automated on KingFisher Flex robots (Thermo Fisher Scientific, USA). Blank controls and mock communities (Zymo Research Corporation, USA) were included and carried through all downstream processing steps. 16S rRNA gene amplification was performed according to the Earth Microbiome Project protocol7. Briefly, Illumina primers with unique forward primer barcodes8 were used to amplify the V4 region of the 16S rRNA gene (515fB-806r9). Amplification was performed as single reactions per sample10, then equal volumes of each amplicon were pooled and the libraries sequenced on the Illumina NovaSeq 6000 sequencing platform with paired-end 150 bp cycles at the Institute for Genomic Medicine (IGM, UC San Diego).