| Extract_protocol | Leaf material were collected from cultivars grown at ENS-Lyon, at Lyon botanical garden, or in a private rosegarden (O. Masquelier, La Bonne Maison, Lyon) (Supplementary Table 4). Approximately 100 mg young leaves were ground in liquid nitrogen using mortar and pestle. No previous nuclei purification step was undertaken, but ground samples were collected in 1.5 ml of homogeneization buffer (Tris 15 mM, EDTA 2 mM, NaCl 20 mM, KCl 80 mM, pH 8.5) with 0.7% (W:V) PVP40, 0.5% (V/V) Triton X100 and 0.1% (V:V) 2-mercaptoethanol. Samples were homogenized for 1h by spinning at 20 cycles / min and pellets were retrieved by 20 min centrifugation at 3000 g. Genomic DNA was then extracted from from pellets using DNeasy Plant kit (Qiagen, MD, USA). DNA integrity was inspected via gel electrophoresis (0.7% agarose) and total DNA was quantified by fluorometry using Picogreen (Applied Biosystems/Life Technologies, Carlsbad CA, USA). |
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