GSM5425864: IR57311 Base HNT; Oryza sativa; RNA-Seq
Homogenized plant material of five replicate samples per cultivar, segment and treatment was pooled to 100 mg resulting into 48 samples for the analysis. Total RNA was isolated using a TRIzol protocol based on the 'single step' method (Sprenger, Kurowsky, Horn, Erban, Seddig, Rudack, Fischer, Walther, Zuther, Köhl, Hincha & Kopka, 2016). Four μg of each RNA sample was further treated with DNase (RapidOut DNA-removal Kit, Thermo Scientific). Absence of genomic DNA contamination was confirmed by qRT-PCR using intron-specific primers (At5g65080). Finally, RNA quality and integrity were verified with Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). Library preperation with NEBNext Ultra™ Directional RNA Library Prep Kit for Illumina (New England Biolabs) and library type RNA library including polyA enrichment. RNA libraries were prepared for sequencing using standard Illumina protocols