GSM2772279: cold-sensitive rice; Oryza sativa Indica Group; RNA-Seq
Total RNA was extracted from rice leaves of nine plants using Concert Plant RNA Reagent (Invitrogen) and treated with DNase I (Invitrogen). Approximately 20 µg of total RNA was used in high-throughput cDNA sequencing by Illumina HiSeq 2000 technology (Fasteris SA, Plan-les-Ouates, Switzerland - http://www.fasteris.com/). We constructed an individual single-end cDNA library for each rice genotype. The cDNA libraries were prepared according to Illumina’s protocols. Briefly, RNAseq was performed using the following successive steps: poly-A purification; cDNA synthesis using a poly-T primer, shotgun method to generate inserts of approximately 500 bp; 3p and 5p adapter ligations; pre-amplification; colony generation; and Illumina single-end 100 bp sequencing.