RNA-seq analysis of strawberry fruit treated hexane
Total RNA was purified using an ISOSPIN Plant RNA kit (Nippon Gene., LTD., Tokyo, Japan) from 10 strawberries treated with hexane 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.
SUB5221432
PR0849_02A04_H1