Qc bioanalyzer

The entire RNA was extracted from T. ammi inflorescences using the TRIzol reagent (Invitrogen, USA) according to the manufacturer’s instruction. The RNA samples were treated with DNase I (TURBO DNase; Ambion, TX, USA). The quality and quantity of the extracted RNA were assessed with 1% agarose gel and NanoDrop 1000 spectrophotometer (Thermo Scientific, USA), respectively. Furthermore, subsequent quality control for the extracted RNA was examined by using a QC Bioanalyzer (Agilent Technologies, Hørsholm, Denmark) and the RNA integrity number (RIN) of each sample was greater than 8. The selection of Poly A, cDNA preparation, adapter ligation, formation of clusters and sequencing was performed at the Beijing Genomes Institute (China), according to the manufacturer’s recommendation, with the use of standard Illumina kits. The sequencing was done on an Illumina HiSeq. 2000 platform with a paired-end and read length of 101 nt.

Total RNA was extracted from 200 mg ground leaves samples (4 leaf stage) using RNeasy Plant Mini Kit (Qiagen, Germany) according to the manufacturer’s protocol. The quantity and quality of the extracted RNA were analyzed using NanoDrop 1000 spectrophotometer (Thermo Scientific, USA) and 1% agarose gel electrophoresis, respectively. The extracted RNA samples were treated with DNase I (Thermo Scientific, USA) to eliminate probable genomic contamination. Each of the control and drought stress treatments had three replications. Subsequent quality control of the extracted RNA was done using a QC Bioanalyzer (Agilent Technologies, CA, USA). The Poly-A selection, preparation of cDNA, adapter ligation, clusters formation, and sequencing was carried out at Beijing Genomes Institute following the manufacturer’s instruction, using TruSeq Stranded total RNA with Ribo-Zero Plant kit (Illumina, USA). The sequencing was performed by Poly-A mRNA Capture method using a Nova-seq 6000 platform to produce 6 GB raw data of 100 bp paired-end reads.