title: RNA-seq analysis of strawberry fruit treated E2-hexenal
design: Total RNA was purified using an ISOSPIN Plant RNA kit (Nippon Gene., LTD., Tokyo, Japan) from 10 strawberries treated with trans-2-hexenal according to the kits protocol. Subsequent processing and RNA-seq analyses were performed using a next-generation sequencer by TaKaRa Bio Inc. (Shiga, Japan). A sequence library was prepared from total RNA using a TruSeq Stranded mRNA Sample Prep Kit (Illumina, Inc. CA, USA), according to the kits preparation guide (Rev. E). PolyA+ RNA was isolated from total RNA and fragmented. Then, single-stranded cDNA was synthesized by reverse transcription using fragmented RNA as a template. Using this single-stranded cDNA as a template, double-stranded cDNA incorporating dUTP was synthesized. After smoothing and phosphorylating both ends of the double-stranded cDNA, 3'-dA was added, and the index adapter was ligated. Adapter-ligated cDNA was amplified by PCR using a polymerase that does not selectively amplify dUTP, and the sequence library was prepared. Library quality was confirmed using an Agilent 2100 Bioanalyzer.