Instrument: Illumina Genome Analyzer IIx
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Three biological replicates were used for all RNA-Seq experiments from each genotypes and water treatment. The total RNA from the leaf meristem was extracted using Trizol reagent (Invitrogen, Carlsbad, CA) and purified using the RNeasy Plant Mini kit (Qiagen, Valencia, CA). On column DNase digestion was performed according to the manufacturer’s protocol (Qiagen, Valencia, CA). RNA quality and integrity was verified using a 2100 Bioanalyzer RNA Nanochip (Agilent, Santa Clara, CA) and all three samples had RNA Integrity Number (RIN) value more than 8.5. The quantification of the total RNA was checked by a NanoDropND-1000 Spectrophotometer (Nano-Drop, Wilmington, DE) and agarose gel electrophoresis. Illumina sequencing using the GAII platform was performed at Beijing Genomics Institute (BGI-Shenzhen, Shenzhen, China http://www.genomics.cn/index.php) according to the manufacturer’s instructions (Illumina, San Diego, CA). Briefly, poly-A RNA was isolated from 20 μg of total RNA using Magnetic Oligo (dT) Beads (Illumina) and digested in short fragment. First and second strand synthesis were followed by end repair, and adenosines were added to the 3’ ends. Adapters were ligated to the cDNA and fragments (200 ± 25 bp) were purified by agarose gel electrophoresis and amplified by PCR. Finally, after validating on an Agilent Technologies 2100 Bioanalyzer using the Agilent DNA 1000 chip kit, the cDNA library was sequenced on a PE flow cell using Illumina Genome AnalyzerIIx, and the workflow was as follows: template hybridization, isothermal amplification, linearization, blocking, sequencing primer hybridization, and sequencing on the sequencer for Read 1