title: RAD_Seq of Lolium multiflorum: NAM founders
design: A segment of 2 cm of mature plant leaf was sampled and stored at -80C for DNA extraction. After grinding the samples with a TissueLyserII (Qiagen, Hilden, Germany) at 25Hz for 40 seconds using two grinding beads (Tungsten Carbide Beads, 3mm, Qiagen, Hilden, Germany), DNA was extracted with the Mag-Bind Plant DNA DS 96 Kit (Omega Bio-tek, Inc. Norcross, GA, USA) according to the recommended protocol by the manufacturer. DNA quality was checked on a 1.2% agarose gel (220V, 300mA, 20min). DNA quantity was measured with Quant-iTPicoGreen ds DNA Assay Kit (ThermoFisher Scientific, Waltham, Massachusetts). The ddRAD libraries were prepared using the restriction enzymes EcoRI and TaqI and followed the protocol (Peterson et al., 2012)