title: GBS EcoT22I koa parents and expected progeny
design: Phyllodes were collected from putative hybrids as well as individual trees and placed on ice in plastic bags. Approximately 130 mg fresh weight of phyllode samples were sprayed with 70% isopropanol, wiped with sterile paper towels, and cut into 2 mm strips for processing. Samples were ground in a sterile mortar and pestle with liquid Nitrogen until they became a fine powder. DNA was extracted using the NucleoSpin Plant II DNA extraction kit (Macherey-Nagel, Bethlehem, PA, USA) following manufacturer s recommendations except as modified by adding an additional centrifugation step and heating of elution buffer. DNA samples were dissolved in TE (10 mM Tris, 0.1 mM EDTA, pH 7.5-8), placed in a 96-well plate, and sent to the Cornell University s Genomic Diversity Facility (Ithaca, NY, USA) for genotyping-by-sequencing. EndonucleaseEcoT22I was used to digest these koa DNA samples.