Transcriptome of rice seeds germinated at ambient and high temperature
Library preparation and Ion Proton sequencing were performed at Genotypic Technology’s Genomics facility following certified protocols from Life Technology.Transcriptome library for sequencing was constructed according to the Ion Total RNA-Seq Kit V2 User Guide (Part # 4476286; Rev. D). The Poly-A RNA was fragmented with RNase III and cleaned using Nucleic Acid Binding Beads provided in Ion Total RNA-Seq KitV2 . The fragmented RNA was hybridized and ligated with adapter and reverse transcribed with ArrayScript Reverse transcriptase. The cDNA was cleaned up using Nucleic Acid Binding Beads provided in Ion Total RNA-Seq Kit V2. The cDNA was subjected to 16cycles of PCR amplification and cleaned using Nucleic Acid Binding Beads provided in Ion Total RNA-Seq Kit V2. The prepared library was quantified using Qubit and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent).Ion Proton sequencing was conducted at Genotypic Technology, Bangalore.