RNA-Seq of Cassia fistula: pods of mature tree, replicate 1
50 mg of pulverized pod tissue were mixed with 1 mL of detergent- based lysis solution (2 % SDS, 0.32 M sodium acetate, 25 mM EDTA), 400 L nuclease- free water, and 100 L acid phenol (pH 4.5). After 5 min at room temperature, 100 L chloroform was added, incubated for 20 min, vortexed, and centrifuged (13,000 rpm, 10 min, 4  C). The aqueous phase was extracted again with acid phenol:chloroform:isoamyl alcohol (25:24:1), and RNA was precipitated by adding 0.6 vol isopropanol (20 C) and 0.4 vol 10 M LiCl. The pellet was washed (70 % ethanol) and resuspended in 30 L of nuclease-free water.