Target Enrichment of Steromapedaliodes albonotata thiemeiana DLC_482_S43
Genomic DNA was extracted from two legs of each individual using the NucleoSpin Tissue Kit (Macherey-Nagel, Germany). DNA was eluted in 100 ul, dd H2O. We used the target enrichment kit for Lepidoptera generated by Mayer et al. (2021), and further molecular protocols follow that study. In short, DNA concentration of samples were measured using a Quantus Fluorometer (Promega), and DNA quality was analysed with a Fragment Analyzer (Advanced Analytical, now Agilent Technologies Inc.). Where necessary, DNA was fragmented using a Bioruptor PICO sonicator (Diagenode). The NEBNext FFPE DNA Repair Mix (NEB, USA) was added to lower quality samples to repair DNA. Reactions were purified using Agencourt AMPure XP beads. End-repair and A-Tailing reactions were conducted using the Agilent Sure Select XT2 Library Prep Kit.Dual-indexed libraries were prepared using the New England Biolabs reaction kits including the NEBNext Quick Ligation Module, adaptors from the NEBNext Multiplex Oligos for Illumina (Dual Index Set1) Kit and the NEBNext Q5 HotStart HiFi PCR Master Mix.Hybridisation was performed with the SureSelect XT2 Pre-Capture ILM Module Box 2 (Agilent Technologies) and the SureSelect Custom Baits 611.9 Mb kit (Agilent Technologies). Samples were sequenced on the Illumina Nextseq 500 System.