ddRADseq of Tetramorium immigrans
the rare EcoRI and common MseI were used for restriction enzyme digestion. Illumina sequencing adaptors and custom barcodes between 8 and 14 bases in length that varied by at least four bases (Parchman et al., 2012) were ligated onto the sticky ends, and PCR amplified using primers for Illumina sequencing adaptors. Amplified digestion-ligation products pooled into equal volumes for sequencing. Gel-based size selection for 200  500bp fragments and sequencing library purification using Agencourt AMPure XP beads (Beckman Coulter) were performed at the UF ICBR core-sequencing laboratory (http://www.biotech.ufl.edu), and sequenced using Illumina NextSeq500 high-output flow cell for a shared 150-cycle sequencing run.