GSM9778491: M2; Oryza sativa; RNA-Seq
Total RNA was extracted from rice leaf tissues using the TRIzol reagent (Invitrogen) following the manufacturer's instructions. RNA integrity and purity were assessed using agarose gel electrophoresis, NanoDrop spectrophotometer and Agilent 2100 Bioanalyzer. Only high-quality RNA samples (RIN ≥ 7.0, 260/280 ratio ≥ 1.8) were used for library construction. Strand-specific RNA-seq libraries were constructed using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer's protocol. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads, fragmented, and reverse transcribed into cDNA. After end repair, A-tailing and adapter ligation, the library fragments were PCR-amplified and purified. Library quality and concentration were verified using the Agilent 2100 Bioanalyzer and qPCR.