2.2 Molecular methods to generate Angiosperms353 sequence data
Total DNA was isolated using a modified CTAB protocol (Doyle & Doyle, 1987). Genomic libraries were constructed as optimized in (Viruelet al. , 2019) using half volumes of the NEBNext® UltraTM II DNA Library Prep Kit for Illumina® (New England Biolabs, Ipswich, MA, United States), purified using AMPure XP magnetic beads and multiplexed with NEBNext® Multiplex Oligos for Illumina® (Dual Index Primer Sets I and II). Equimolar pools containing twelve genomic libraries were enriched with half-reactions of the Angiosperms353 probe kit (Johnson et al. , 2019; Baker et al. , 2022) following myBaits®kit manual v5.03 (Arbor Biosciences). DNA concentrations were calculated using a QuantusTM fluorometer (Promega Corp.), and an Agilent 4200 TapeStation (Agilent Technologies, Santa Clara, CA, United States) was used to assess fragment length. Sequencing was performed on a HiSeq (Illumina, Inc.) by Macrogen (Seoul, South Korea), producing 150 bp paired-end reads.