RNA-Seq of Arabidopsis shoot: Plasma-Activated Water -3
Three-day old Col-0 Arabidopsis seedlings were transferred onto gel growth media containing PAW4 or the equivalent NO3- control and grown for additional 8 days. The shoot and root tissue of these seedlings were excised, and excess gel media was removed immediately through gentle dabbing. Samples were pooled into four replicates of approximately 20 Arabidopsis seedlings each and placed in 5 volumes of RNAlater stabilization solution. Seedlings spent less than 10 seconds from harvesting to transfer to RNAlater. RNA was then extracted from the tissue using the RNeasy plant mini kit (Qiagen, 74904). Quality of extracted RNA was tested utilizing an Agilent 4200 Tapestation. cDNA library preparation was performed using the NEB NEBNext Ultra II Direction RNA kit (polyA enriched) (NEB, E7760S). Next-Generation Sequencing was conducted using an Illumina NovaSeq 6000 with 150-base paired-end reads.