title: GSM8417227: Mild-drought_fixed_rep1; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: The cell wall digestion protocol was adapted fromRyu et al. (2019). Briefly, the third leaf of 80 plants per sample was harvested, chopped in 1 mL enzyme solution (0.4 M mannitol, 10 mM CaCl2, 20 mM MES, 20 mM KCl, 2% (wt/vol, 5.7 pH) cellulase, and 0.5% (wt/vol) macero enzyme). Chopped samples were transferred to a 70-μm cell strainer (model Corning, Merck) placed within a 6-well plate, in a total volume of 5  mL of the enzyme solution. In the fixed samples, 50uM of Actinomycin D (Sigma A9415) was added to the cell wall digestion solution. Samples were incubated, constantly shaking for 75 min at room temperature. After a 7-min centrifugation at 200g, the pellet was resuspended in 1 mL washing solution (0.4 M mannitol, 10 mM CaCl2, 20 mM MES, and 20 mM KCl). Cells were sorted on the LeviCell magnetic sorter. Subsequently, cells were collected in wash buffer (0.4 M mannitol, 10 mM CaCl2, 20 mM MES, and 20 mM KCl) and counted. Cellular suspensions were loaded into a BD Rhapsody HT Cartridge (BD) according to the manufacturer's instructions.