Sorghum bicolor 7
Total RNA was extracted using miRNeasy Mini Kit (Qiagen, USA#217004) and treated with DNase I using in column DNase I digestion as described in the users manual. The integrity of RNA was checked using Bioanalyzer (Agilent Technologies) and RNA samples with RNA integrity number (RIN) ~ 9.0 or more were used for further analyses. Poly (A) mRNA enrichment and construction of stranded mRNA-seq libraries were performed using KAPA Stranded mRNA-seq kit (Cat# KR0960) (Illumina). Briefly, poly (A) mRNA was captured using magnetic oligo (dT) beads and fragmented using heat and magnesium. First-strand cDNA was synthesized using random primer and the second strand was synthesized in the presence of dUTP. Adenosine was added to the 3-end followed by adaptor ligation. The library was amplified using high fidelity, low-bias PCR. Single-end sequencing of the libraries was done at the Sequencing & Analysis Core Resource, Duke University using Illumina Hi seq 2000.