UCE sequences of adult Schwarzia icipensis sp. nov.
We generally soaked single whole specimens in Proteinase-K extraction buffer from DNeasy Blood and Tissue Kits (Qiagen Inc.) and incubated overnight at 56C (~12 h), or used a CTAB phenol-chloroform protocol for extraction. After incubation, whole specimens were washed in ethanol and mounted back on pins. For the single specimen of S. lualenyiensis, which represents the holotype for the new species, we carefully broke off a single hindleg and glued it back on to the specimen after incubation. We followed the extraction kit manufacturers protocol but eluted the DNA from the spin column with 120 l of purified H2O. We assessed DNA fragmentation and concentration, and sheared fragments to a targeted size of 450-600 bp using a Q800R sonicator (Qsonica, LLC.). We used up to 100 ng of DNA per sample to prepare individual dual-indexed libraries with a KAPA HyperPrep kit (Roche Sequencing Systems, Inc.) and TruSeq adapters (Faircloth and Glenn 2012). We followed the pooled on-bead enrichment procedure outlined in Faircloth et al. (2015) but included the modifications from Blaimer et al. (2016b, 2016a). We used the expanded Hymenoptera UCE bait set (principal Hym-v2; Branstetter et al. 2017) or the bee-ant-specific hym-v2 bait set from Grab et al. (2019); both manufactured by MYcroarray, Inc (or Arbor Biosciences). Following post-enrichment amplifications, we measured DNA quantity with a Qubit fluorometer (model 3.0, Thermo Fisher Scientific, Inc.) and assessed fragment distribution with an Agilent 2200 TapeStation and D1000 HS screen tapes (Agilent Technologies, Inc.). Based on our results, we combined pools at equimolar concentrations and size selected for a range of 250-600 bp with a Blue Pippin device (Sage Science, Inc.). Sequencing was carried out on Illumina HiSeq platforms with 150 bp read lengths. The adapter combination for this species samples is: ACGTTACC+GCTTCGAA.