title: 16s sequencing of Trachymyrmex arizonensis Clade-A fungus microbiome
design: DNA extraction and sequencing of the 16S samples was performed at MRDNA in Shallowater, Texas (http://www.mrdnalab.com/). DNA sequences were amplified from whole ants and fungus using primers 27F 5' AGRGTTTGATCMTGGCTCAG and 519R 5' GTNTTACNGCGGCKGCTG that span the V1-V3 hypervariable regions of the 16S rRNA gene. They were processed using the HotStarTaq Plus Master Mix Kit (Qiagen, USA) under the following conditions: 94C for 3 minutes, followed by 28 cycles of 94C for 30 seconds, 53C for 40 seconds and 72C for 1 minute, after which a final elongation step at 72C for 5 minutes was performed. After the samples were amplified and checked for adequate genetic yields, the sub-samples were pooled back together and purified using calibrated Ampure XP beads. The purified and pooled PCR product was used to create a DNA library and sequenced using the Illumina MiSeq platform in PEx300 mode