RNA-Seq of glycine max: whole mature seed
Seed samples collected in the treatments were frozen in liquid nitrogen (N2) and stored at -80 C until RNA extraction. Samples were macerated in liquid N2, and total RNA was extracted using the Concert Plant RNA reagent kit (Invitrogen, California, USA) according to the manufacturer's specifications. RNA quantification was performed in a NanoDrop spectrophotometer according to the following quality and purity parameters: concentration > 600 ng L-1, 260/280 ratio ranging from 1.8 to 2.0, and 260/230 ratio  2.0.The RNA was treated with the DNAse-free turbo kit to remove the remaining ge-nomic DNA (Invitrogen, California, USA). RNA integrity was assessed by electrophoresis on a 1% (w/v) agarose gel stained with ethidium bromide (1 g mL-1) (Sambrook et al., 1989). High-quality total RNA samples were sent for sequencing at the University of Georgia (Georgia Genomics Facility  GGF, USA). Before the samples were sequenced, they were evaluated on an Agilent Bioanalyzer 2100 instrument (Agilent Technologies, Inc.). Only samples with an RNA integrity number (RIN)  7.00 were used to synthesize the mRNA-Seq libraries. Libraries were synthesized using the Ilumina TruSeq SBS v5 poly-A kit on an Illumina NextSeq 500 1.9 75 bp paired-end sequencing platform (Illumina, San Diego, CA, USA), with at least 1X genome coverage.