title: BH5_23C_BTH_2
design: Four-week-old Arabidopsis thaliana accession Columbia-0 plants were used for treatments.   Plants were moved from 23C chambers to 30C chambers 2 h after lights on and acclimated for 24 h.  Plants were then sprayed with either a 0.1% dimethyl sulfoxide (DMSO), 0.01% Silwet L-77 (OSi Specialties) mock solution or a 100 然 benzo(1,2,3)thiadiazole-7-carbothioic acid-S-methyl ester (BTH; Chem Service Inc., Cat# N-11026) in 0.1% DMSO, 0.01% Silwet solution and returned to the either 23C or 30C chambers.  Twenty four hours later, two fully expanded leaves were harvested and flash frozen in liquid nitrogen.  RNA was extracted from flash-frozen, ground leaf tissue following the manufacturer掇 protocol (Ambion ToTally RNA kit).  Samples were digested with DNaseI (Roche) to remove any genomic DNA contamination, and then purified using the RNeasy Mini kit (Qiagen).  Sequencing libraries were prepared from oligo-dT purified RNA using the Illumina TruSeq library preparation protocol.  Sequencing was performed in a 1x50bp single end format using HiSeq Rapid SBS reagents. Base calling was done by Illumina Real Time Analysis (RTA) V1.18.64 and output of RTA was demultiplexed and converted to FastQ format with Illumina Bcl2fastq v1.8.4.