lcWGS of costus genetic mapping population: sample 173
To identify QTL underlying floral trait variation associated with bee-to-hummingbird pollination shifts, low-coverage whole genome sequencing data was generating for a mapping population of Costus bracteatus x Costus lasius F2 hybrids. Either young leaf tissue or meristem tissue was collected from all F2 hybrids and dried in silica gel prior to extraction with the NucleoSpin Plant Mini Kit II (Macherey-Nagel, Duren, Germany) according to the manufacturers protocol, adding 5 L proteinase K (20 mg/mL) to the digestion step and increasing the digestion incubation time to an hour. Genomic DNA was quantified with a Qubit dsDNA BR assay (Qiagen Inc., Valencia, CA, USA) and quality-checked with Nanodrop and gel electrophoresis. Whole-genome shotgun libraries were prepared, sequenced, and demultiplexed by the University of California Davis DNA Technologies and Expression Analysis Core (Davis, CA). For each sample, 10 ng of genomic DNA was converted to barcode-indexed sequencing libraries using the Seqwell plexWell LP384 Library Preparation Kit for Illumina Sequencing Platforms (seqWell, Beverly, MA). The libraries were amplified with eight PCR cycles and analyzed with a Bioanalyzer 2100 instrument (Agilent, Santa Clara, CA), quantified by fluorometry on a Qubit instrument (LifeTechnologies, Carlsbad, CA), and combined in two pools at equimolar ratios. The library pool was quantified by qPCR with a Kapa Library-Quant kit (Kapa Biosystems/Roche, Basel Switzerland) and sequenced on an Illumina NovaSeq system (Illumina, San Diego, CA) run with paired-end 150 bp reads.