GSM5706340: NIP_6h_3; Oryza sativa Japonica Group; RNA-Seq
Every sample was pooled with infected leaves collected from 15 plants in one plot to diminish the individual environment differences.Total RNA from the mentioned samples were extracted using Trizol reagent (Invitrogen, USA) according to the manufacturer's recommendation. NEB Next Poly(A) mRNA Magnetic Isolation Module (NEB) kit was used to enrich the poly(A) tailed mRNA molecules from 1 μg total RNA. The mRNA was fragmented into ~200 base pair pieces. The first-strand cDNA was synthesized from the mRNA fragments reverse transcriptase and random hexamer primers, and then the second-strand cDNA was synthesized using DNA  polymerase I and RNaseH. The end of the cDNA fragment was subjected to an end repair process that included the addition of a single “A” base, followed by ligation of the adapters. Products were purified and enriched by polymerase chain reaction (PCR) to amplify the library DNA. The final libraries were quantified using KAPA Library Quantification kit (KAPA Biosystems, South Africa) and an Agilent 2100 Bioanalyzer. After quantitative reverse transcription-polymerase chain reaction (RT-qPCR) validation, libraries were subjected to paired-end sequencing with pair  end 150-base pair reading length on an Illumina NovaSeq sequencer (Illumina)