16S metagenome of Xyleborus affinis collected from a Bursera simaruba infested branch
Ribosomal amplicons of the variable V3 region of the 16S gene was amplified from five nanograms per microliter of total DNA extracted from the Xyleborus affinis collected from a Bursera simaruba infested branch. 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).