16S metagenome from the abdomen of Xyleborus affinis reared with Persea schiedeana
Ribosomal amplicons of the variable V3 region of the 16S gene was amplified from five nanograms per microliter of total DNA extracted from the abdomen of Xyleborus affinis reared on an artificial diet prepared with Coyo Avocado (Persea schiedeana) sawdust. Illumina libraries were constructed by adding Nextera XT adapters (Illumina Inc., U.S.A.). Amplicon PCR and adapter-ligation PCR were performed using Kapa polymerase (Kappa HiFi Hotstart Ready Mix, cat. No. KK2602, Kappa Biosystems) following the manufacturers recommendations and purified immediately after finishing each PCR using 0.8X Agencourt Ampure XP cleaning beads (BeckmanCoulter, cat. No. A63881). Library concentration was quantified using a DNA HS kit (Invitrogen, cat. No. Q32854) in a Qubit 2.0 fluorometer (Invitrogen, cat. No. Q32866). Adapter ligation and library size were evaluated using a Bioanalyzer (Agilent, cat. No. G2953CA) DNA HS (Agilent, cat. No. 5067-4626) chip. Libraries were diluted and pooled to an equimolar concentration to be denatured and loaded in a MiSeq (Illumina Inc., U.S.A.) using a MiSeq Reagent kit V3 (Illumina Inc. cat. No. 15067874).