title: ITS2 community amplicon-seq of Trachymyrmex septentrionalis fungus garden: field-collection FL, USA
design: Fungus garden samples stored in DESS were rinsed with 1 X PBS three times before DNA was extracted using a modified CTAB protocol. DNA was amplified for the ITS2 region using primers fITS7 and ITS4 and then cleaned using QIAGEN QIAquick Gel Extraction Kit. Libraries were prepared using EpMotion 5075. Each sample was amplified in triplicate using Accuprime buffer/polymerase and sequencing primers fITS2 & ITS4, each with barcodes and Illumina adapters. Sample triplicates were re-pooled, checked for amplification, and then all barcoded samples were pooled into one library. Libary concentraion was adjusted using Qubit with dsHS DNA kit. Library was sequenced on Illumina MiSeq using 2x250 chemistry.