Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA was isolated using the TRIzol reagent (Invitrogen Life Technologies, USA) following the manufacturer's instructions and was treated with RNase-free DNase I (Takara, Japan) to avoid genomic DNA contamination.For each time point, RNA was extracted from 6 randomly selected seedlings (as one biological replicate). Two replicates were conducted for RNA extraction for each time point. The poly (A) mRNA was isolated from the total RNA samples using oligo (dT) attached magnetic beads (Invitrogen) and was fragmented into short pieces using divalent cations under elevated temperature. First- and second-strand cDNA was generated using mRNA-Seq sample preparation (Illumina Inc., San Diego, CA, USA) and random hexamer primers. The double-stranded cDNA fragments were subjected to end repair, and sequencing adapters were ligated to both ends. The final cDNA library was selectively enriched by PCR and purified using the AMPure XP system (Beckman Coulter, Beverly, USA).