Genomic DNA (gDNA) was extracted from fresh frozen colorectal cancer tissue with the E.Z.N.A.® tissue DNA Kit (Omega Bio-tek, U.S.) and from fecal samples with the E.Z.N.A.® soil DNA Kit (Omega Bio-tek, U.S.) according to manufacturer’s instructions. The V3-V4 regions of the 16S rRNA gene were PCR amplified using primer pairs 338F (5′-ACTCCTACGGGAGGCAGCAG-3′) and 806R (5′ GGACTACHVGGGTWTCTAAT-3′). The PCR reaction was set up in triplicate, and the PCR product was purified using the AxyPrep DNA Gel Extraction Kit (Axygen Biosciences, USA). The high-throughput sequencing was performed in Majorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China). Analysis of the 16S rRNA microbiome sequencing data was performed using the free online platform of Majorbio Cloud Platform (cloud.majorbio.com).
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