Identification of the FRS12 regulated genes by RNA-Seq Analysis in Arabidopsis thaliana

To identify genes regulated by FRS12 at night time, stratified Arabidopsis thaliana transgenic seedlings expressing Pro35S:FRS12-GR and Pro35S:GFP-GR were grown in 1/2 MS with 1% sucrose liquid medium under long days (16:8) conditions and treated with 5uM of dexametasone 4H before harvesting at night time at zeitgeber time 23 (time of lights on usually defines zeitgeber time zero). Three biological replicates were harvested for each line. RNA was isolated using the RNeasy Plant Mini Kit (Qiagen) and DNase I treated (Promega). RNA samples were processed by first preparing a TruSeq RNA-Seq library (Illumina) and sequenced at 50bp single read using Illumina HiSeq 2000.