RNA-Seq of Glycine max: imbibed germinating embryonic axis from 3-year-old seed
Soybean seeds harvested in 2015 (aged 3 years in dry storage),  1999 (19 years), and 1996 (22 years) were imbibed for 24 hours in the dark. Embryonic axes was removed and individually flash-frozen in liquid nitrogen. The associated cotyledons remained in the light for 72 hours and were then assessed for greening  - seeds that greened were likely to have germinated and seeds that failed to green were not. RNA was extracted from imbibed as well as dry axes from all three harvest years with the Qiagen RNeasy kit and poly-A selection was used to isolate mRNA followed by cDNA synthesis. Individual axes were sequenced.