title: ITS3-ITS4 Fluidigm Pool
design: All DNA samples were initially quantified using the Qubit Fluorometer 2.0 (Thermo Fisher Scientific). Polymerase Chain Reaction (PCR) amplification was performed using three primer sets (as found in the metadata): two primer sets for amplification of two variable regions (V3-V5 and V4) of the bacterial 16S ribosomal RNA (rRNA), and a primer set for amplification of a portion of the nuclear ribosomal Internal Transcribed Spacer (ITS) region (ITS 3-4) in fungi. A 48.48 Fluidigm Access Array (Fluidigm Corporation, South San Francisco, USA) was used for amplification and library construction.  PCR products were harvested from the PCR chambers from the Post-PCR IFC Controller AX.  Amplicons containing sample-specific barcodes and Illumina adapter sequencers were quantified with a Fragment Analyzer Automated CE System (Advanced Analytical) and pooled in equimolar ratios into two Eppendorf tubes; DNA bands in the size range of 350-1000 bp were selected and excised following gel electrophoresis corresponding to equal amplicon size of V3-V5, V4 and ITS3-4 products. DNA was purified with a Mini Elute Gel Extraction Kit (Qiagen), then analyzed and quantitated.