UCE target enrichment of Eleutheronema_tetradactylum_L
We input 110-1,100 ng into Kapa Biosystems DNA library preparation kit that incorporated “with-bead” cleanup steps during end-repair, +A addition, and ligation steps.  We substituted adapters (12 µM) containing a custom index sequence (Faircloth and Glenn 2012) for the standard Illumina adapters, and we used a generic SPRI substitute (Rohland and Reich 2012; hereafter SPRI) in place of commercially available AMPure reagent (Beckman-Coulter) for all AMPure cleanup steps. We amplified 15 µL of the library volume using a reaction mix of 25 µL HiFi HotStart ReadyMix (Kapa Biosystems), 5 µL of Illumina TruSeq primer mix (5 µM each), and 5 µL ddH20 and the following thermal profile: 98 °C for 45s; 10-12 cycles of 98 °C for 15s, 60 °C for 30s, 72 °C for 60s; and a final extension of 72 °C for 5m. We combined groups of eight libraries at equimolar ratios into enrichment pools having a final concentration of 147 ng/µL in 3.4 µL (500 ng total). We enriched libraries using an acanthomorph probe set targeting 1314 UCE loci and containing 2628 probes (Alfao et al. in prep), and we followed library enrichment procedures for the MYcroarray MYBaits kit
SUB1977116
Eleutheronema_tetradactylum_L