RNAseq of Arabidopsis seeds during hydropriming
RNA was isolated from 450 mg dry material with 10 ml of polysome extraction buffer. The aliquots of total and ribosome associated RNA were further prepared for RNA sequencing using clean-up RNeasy Mini Kit (Qiagen, Hilden, Germany). All RNA samples were sequenced with the Illumina NovaSeq 6000 platform (Illumina, Inc, San Diego, CA, US), following the PE150 strategy for unstranded sequencing. For this, the RNA samples were first fragmented into pieces of approximately 250 bp. RNA fragments were reverse transcribed to cDNA with random primers after which the cDNAs were dA-tailed. The dA-tailed DNA fragments were then ligated tosequencing adaptors. The final DNA library was obtained with PCR amplification