SC216-2
The total RNA in each sample was extracted from the leaf tissues samples using TRIzol reagent following the manufacturers instructions (Invitrogen, Carlsbad, California, USA), and genomic DNA was removed using DNase I (TaKaRa Bio, Mountain View, California, USA). RNA quality was checked by Agilent bioanalyzer 2100 (Agilent Technologies, Santa Clara, California, USA), and Nanodrop 2000 spectrophotometer (Nanodrop Technologies, Wilmington, Delaware, USA) was used for RNA quantification. RNA-seq libraries construction of the qualified samples was conducted following the TruSeqTM stranded mRNA sample preparation kit (Illumina, San Diego, California, USA). A sequencing library was prepared by random fragmentation of the cDNA samples, followed by 5 and 3 Illumina TruSeq3 adapter ligation. Adapter-ligated fragments were PCR-amplified and gel-purified before being loaded onto a flow cell where fragments were captured on a lawn of surface-bound oligos complementary to the library adapters. Each fragment was then amplified into distinct, clonal clusters through bridge amplification. Paired-end libraries were sequenced from the clusters generated by an Illumina Hiseq X Ten platform (2101 bp read length). Base calling was performed by the Illumina sequencer generated raw images through an integrated primary analysis software package called RTA (Real Time Analysis) and base calls binary was converted into FASTQ files using Illumina package bcl2fastq.