GSM5239810: 2MWT-stage1-rep3; Glycine max; RNA-Seq
Once the appropriate stage was reached, seed coats and radicles were removed, and the tissue was flash frozen with liquid nitrogen and stored at -80°C until use. The tissue from these stages was ground to powder with mortar, pestle, and liquid nitrogen. Total RNA from all stages was extracted using the RNeasy Plant Kit with on-column DNase digestion and RLC buffer (QIAGEN, Hilden, Germany). RNA quality was determined by UV spectrophotometry (260 nm, NanoDrop1000, Thermo Fischer Scientific, Waltham, MA) and RNA integrity numbers (RIN) (BioAnalyzer, Agilent Technologies, Santa Clara, CA). Samples with a RIN value >8.0 and 260/280 ratios >2.0 were submitted to Novogene (Sacramento, CA) for mRNA sequencing. A total of 72 samples (8 lines x 3 germination stages x 3 biological replicates) were sequenced (Illumina, San Diego, CA) to acquire 30 million, 150 PE reads per sample. paired end