Characterization of the endophyte mycobiome in Cowpea (Vigna unguiculata) using an Illumina Sequencing Approach
Metagenomic DNA from 0.1g of each pulverized plant sample was extracted using the Nucleospin Plant II mini Kit (Macherey Nagel, Germany). DNA concentrations were determined using a Nanodrop LITE spectrophotometer (Thermo Scientific, USA) and diluted to a standargd 10 ng/L for Polymerase Chain Reactions (PCR).  The Internal Transcribed Spacer region of the ribosomal operon was amplified using the primers ITS3F (5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGCATCGATGAAGAACGCAGC3) and ITS4R (5-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGTCCTCCGCTTATTGATATG-3), with Illumina MiSeq (Sigma Aldrich, USA) specific adaptor sequences (Caporaso et al. 2012).