RNA-Seq library preparation and Illumina sequencing

The libraries were created with high-quality total RNA samples (OD_260/280 = 1.8 ~ 2.2; OD_260/230 ≥ 2.0; RNA Integrity Number (RIN) ≥ 8 out of 10; 28S:18S > 1.0). The cDNA libraries were prepared according to Illumina’s protocol (Illumina Inc., San Diego, USA; cat. no. RS-100-0801) by a genome sequencing company (Novo gene, China) and were sequenced to a depth of 40 million paired-end reads per each biological replicate using an Illumina HiSeq™ 2500 platform (150 bp; Illumina).

The transcriptome raw sequencing data of S. khuzistanica and S. rechingeri have been submitted to the NCBI GenBank (https://www.ncbi.nlm.nih.gov) with BioSample accession numbers as following: SAMN09079833, SAMN09079834, SAMN09079835 and SAMN09080197, SAMN09080198, SAMN09080199 and BioProject accession numbers: PRJNA464236, PRJNA464241.