Colon Cancer Tissue Microbiome
Metagenomic 16S amplicon sequencing was carried out by amplifying the V3-V4 regions of 16S rRNA using primers with Illumina adapter sequences. PCR reactions used 12.5 ng of DNA, KAPA HiFi HotStart Ready Mix, and specific primers, followed by a second index PCR to add barcodes and adapters. The libraries were cleaned, normalized, pooled, and sequenced on the MiSeq with a 600-cycle paired-end run.