GSM7908357: Root tip, lbd quad 16h post cut, Rep 1; Arabidopsis thaliana; RNA-Seq
Primary roots tip (~1cm) were collected into a 35X10mm petri dish containing 60µl of cell wall digestion solution (3% w/v Cellulase, 1%w/v Macerozyme (Yakult), 0.4M Mannitol, 20mM MES, 0.02M KCl, Tris-HCl pH=5.7, heated to 55C° for 10min, cooled to room temperature followed by addition of 0.001 gr/ml BSA and 20mM CaCl2). Meristems were collected after 15min incubation and transferred to a fresh digestion solution for 1h. The solution was filtered twice through 40µm cell strainers and centrifuged for 10 minutes at 300g. Cells pellet were resuspended in washing solution (0.4 M mannitol, 20 mM MES, 20 mM KCl and 0.1% BSA) to a concentration of ~1000 cells/µl Library was prepared according to the manufactorer instructions (10x genomics chromium 3' v2 protocol)