| Description |
The plant tissue samples were frozen in liquid nitrogen, quenched using a Mixer Mill (MM300, Retsch, Hann, Germany) as previously reported ( Hirai et al. 2007 ), and the extraction buffer (80% MeOH) was added. The resulting extracts consisted of 400 μ l with 20 mg ml –1 (fresh weight) of tissue. The extracts were transferred and treated using an ALHS as follows (Supplementary Fig. S1): 350 μ l of each plant extract were transferred to a 96-well plate, and the solutions were dried under N 2 gas using a 96-well format spray instrument (40°C, 25 min and 30°C, 20 min). The dried samples were dissolved in 350 μ l of H 2 O using a vortex system (1,300 r.p.m., 6 min), and then fi ltered through a 96-well filter [Captiva 96-well Filter Plate (pore size 0.45 μ m, polyvinylidene fl uoride), Varian, CA, USA], using a vacuum manifold with the following program: 30 s, 50 Hpa; 20 s, 100 Hpa; 20 s, 200 Hpa; 30 s, 300 Hpa; and 60 s, 300 Hpa. Plate handling was carried out automatically using robot arms (iSWAP, Hamilton). |