18S metagenome of Xyleborus volvulus collected from a Persea americana infested branch
Ribosomal amplicons of an 18S region were prepared to identify fungal diversity using the primers SSUfungiF 5-TGGAGGGCAAGTCTGGTG-3 and SSUFungiR 5-TCGGCATAGTTTATGGTTAAG-3 from five nanograms per microliter of total DNA extracted from the Xyleborus volvulus collected from a Persea americana 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).