Punam_Smear_mucus
Library Preparation and Sequencing. The libraries were prepared following the instructions of the 16S Metagenomic Sequencing Library Preparation for Illumina. The amplicon concentration was assessed using a Qubit 2.0 fluorometer with the dsDNA high sensitivity kit (Thermo Scientific, USA). Amplicon clean-up was carried out using AMPure XP beads (Beckman Coulter). Cleaned amplicons were indexed with the Illumina sequencing adapters using the Nextera XT Index Kit. The indexed amplicons were purified using AMPure XP beads and quantified fluorometrically with the dsDNA high-sensitivity kit (Thermo Scientific, USA). The integrity and quality of the final library were verified using the Agilent High Sensitivity DNA kit on the Agilent Bioanalyzer 2100 system (Agilent Technologies, USA). Libraries were pooled in equimolar concentrations and then sequenced in a paired-end (2  300 bp) sequencing format with a MiSeq Reagent Kit V3 (600 cycles) using the MiSeq platform (Illumina, San Diego, CA, USA). Sequencing was performed in the Sequencing Laboratory at the Instituto de Ecologa, A.C. (INECOL), Mexico.