Hyb-Seq of Myrtaceae: Eugenia excelsa
Sequencing libraries were created with the NEBNext Ultra kit (New England Biolabs), using 200 ng of fragmented DNA for each sample and size-selecting at 550 bp with Ampure magnetic beads. Libraries were prepared in 25 L (half volume) to maximize reagents and indexes, then amplified through eight cycles of PCR amplification. DNA was quantified using a Quantus fluorometer (Promega Corporation, Madison, Wisconsin, USA) and fragment size was estimated using the TapeStation. Dual indexed samples were pooled with up to 50 ng of library DNA for enrichment using the Angiosperm-353 v1 target capture kit (Johnson et al., 2019) available from Arbor Biosciences (Ann Arbor, Michigan, USA), performed over 24 h at 65 C. Enriched products were PCR-amplified for 12 cycles and sequenced on an Illumina MiSeq (Illumina San-Diego, California, USA) with v3 reagent chemistry (2 x 300 bp paired-end reads)