title: GSM1180963: Foc1 3h; Musa acuminata; RNA-Seq
library_construction_protocol: Total RNA was extracted from roots, pseudostems, leaves, floral organs, and developing fruits separately using a modified CTAB method briefly described below. Two to five grams of tissues were grounded in liquid nitrogen, and the powder was mixed with 20 mL CTAB buffer (3% CTAB, 2% PVP, 2.0 M NaCl, 20 mM EDTA, 100 mM Tris pH 8.0, 2% β- mercaptoethanol) and incubated at 65ºC for 20 min. The extract was mixed with 0.6 volume of chloroform by vortexing and span at 12000g for 15 min at room temperature. The supernatant was transferred to a new tube and extracted with an equal volume of chloroform, and the supernatant was then mixed with 0.5 volume of 12 M LiCl and incubated at -20ºC for 2 hours. RNA was precipitated by centrifugation at 12000g for 15 min at 4ºC and the pellet was re-suspended in 1 mL 0.2 M NaCl. The RNA solution was extracted sequentially with an equal volume of water-saturated phenol (pH 4.2) and chloroform. RNA was precipitated by mixing the solution with three volumes of ethanol and leaving on ice for 30 min before centrifugation at 14000g for 20 min at 4ºC. After washing the pellet with 75% ethanol, the RNA pellet was dissolved in 50 μL RNase-free water. RNA libraries were prepared for sequencing using standard Illumina protocols