title: GSM3381668: ir29c-ps; Oryza sativa; OTHER
library_construction_protocol: RNA was isolated from 10 µg each of the total RNA and polysomal RNA using MicroPoly(A) Purist ™ kit (Ambion). Briefly, rice seedlings were ground into a fine powder with liquid nitrogen, then thoroughly mixed with polysome extraction buffer using a spatula and placed on ice for 10 min with occasional mixing by inverting tubes. The lysis was spun for 2 min at 14000 rpm (4  C) and the supernatant was passed through a QIA shredder (QIAGEN) column by centrifugation for 1 min at 14000 rpm (4  C). Approximately 600   microL of the sample was layered on top of the sucrose density gradients (20-60% sucrose, w/v) and centrifuged for 120 min at 40,000 rpm (275,000    g) in a Beckman OPTIMA LE-80 centrifuge. The optical density (OD) of the samples was measured by using a UA-5 detector and a Gradient Fractionator (model 640, ISCO) reading absorbance throughout the sucrose gradient at 254 nm. Fractions with more than two ribosomes were pooled and used for RNA isolation and RNA-Seq library preparation later. mRNA-Seq libraries were generated by following Illumina mRNA Sequencing Sample Preparation Guide.
identifiers.namespace: GEO
identifiers.id: GSM3381668