UCE sequences of Sericomyrmex_amabilis_1642_PA
DNA was extracted destructively or non-destructively from worker, male or pupae ants using a DNeasy Blood and TissueKit (Qiagen, CA, USA). We quantified DNA for each sample with Qubit fluorometer (Life Technologies, Inc.) and sheared to a target size of approximately 500-600 bp by sonication (Q800; Qsonica Inc.). The sheared DNA was used as input for a modified genomic DNA library preparation protocol (Kapa Hyper Prep Library Kit, Kapa Biosystems) that incorporated Ňwith-beadÓ cleanup steps (Fisher et al. 2011) and a generic SPRI substitute (Rohland and Reich 2012, speedbeads), as described by (Faircloth et al. 2015). We used TruSeq-style adapters during adapter ligation (Faircloth and Glenn 2012), and PCR amplified 50% of the resulting library volume (15 uL) using a reaction mix of 25 _L HiFi HotStart polymerase (Kapa Biosystems), 2.5 uL each of Illumina TruSeq-style i5 and i7 primers  and 5 uL double-distilled water (ddH20). We used the following thermal protocol: 98 C for 45 s; 13 cycles of 98 C for 15 s, 65 _C for 30 s, 72 C for 60 s, and final extension at 72 C for 5 m. After rehydrating in 23uL Elution Buffer (EB) and purifying reactions using speedbeads, we combined groups of eight libraries at equimolar ratios into enrichment pools. This final pool was then size-selected to a fragment range of 250Đ800 bp using the BluePippin size-selection instrument (SageScience, Beverly, MA). Size-selected library pools were sequenced as 3 partial lanes (each of the lanes included other samples) on an Illumina HiSeq 2500 (2x150 rapid run).
SUB2101092
1642