title: 16S Amplicon of rectal cancer patients
design: Fifty nanograms of genomic DNA from all fresh-frozen biopsies were used to generate amplicons to evaluate the microbiota (16S rRNA V3-V4 region). Library triplicates were purified with Ampure XP Beads (Beckman Coulter, USA) and pooled, followed by a quantification step by real time PCR (KAPA Library Quantification Kit for Illumina Platforms  KAPA Biosystems, Sigma Aldrich, USA). Sequencing was performed in the MiSeq platform (Illumina, USA) using MiSeq Reagent v2 (500-cycles) in paired-end mode.