P88-3, PI 88788 roots replicate 3
Glycine max
Protocols: The soybean seeds were pre-germinated on moistened filter paper in a plant growth chamber at 27 °C, 85% ambient humidity and a 16:8 (light: dark) photoperiod for 3-4 days. The seedlings were then transferred into tubes (5 cm 3 30 cm) with sandy soil at 27°C under 16 h of light in the greenhouse. When the plants had three fully expanded trifoliate leaves, roots were harvested, flash frozen in liquid nitrogen, ground to powder, and stored at -80 °C until the next step. For each sample, three biological replicates were set up. The soybean seeds were pre-germinated on moistened filter paper in a plant growth chamber at 27 °C, 85% ambient humidity and a 16:8 (light: dark) photoperiod for 3-4 days. The seedlings were then transferred into tubes (5 cm 3 30 cm) with sandy soil at 27°C under 16 h of light in the greenhouse. The root samples were ground into two parts, one of which was used to extract total RNAs using TRIZOL reagent (Invitrogen, CA, USA) following the manufacturer's procedure. The total RNA concentration and purity were assayed with a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). The RNA integrity was assessed on an Agilent 2100 Bioanalyzer Lab-on-Chip system (Agilent Technologies, Palo Alto, CA, USA). The construction of cDNA libraries was according to the protocol for the mRNA-Seq sample preparation kit (Illumina, San Diego, USA). The clustering of samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia, San Diego, USA) according to the manufacturer's instructions.