Whole transcriptome analysis kit

Using the TRIzol Reagent (Catalog No. 15596–026, Invitrogen), total RNAs were extracted from seedlings without or with salt stress treatment. Polyadenylated RNAs were isolated using the Oligotex mRNA Midi Kit (Catalog No. 70042, Qiagen). The RNA-seq libraries were constructed using the Illumina Whole Transcriptome Analysis Kit following the standard protocol (Illumina, HiSeq system) and sequenced by the Bioscience Core Facility at KAUST on the HiSeq platform to generate high-quality pair-end reads.

Using the TRIzol Reagent (15596–026, Invitrogen Co., Carlsbad, CA, USA), total RNAs were extracted from 12-day-old seedlings of wild-type C24, sad1 and SAD1-OE. Polyadenylated RNAs were isolated using the Oligotex mRNA Midi Kit (70042, Qiagen Inc., Valencia, CA, USA). The RNA-seq libraries were constructed using Illumina Whole Transcriptome Analysis Kit following the standard protocol (Illumina, HiSeq system) and sequenced on the HiSeq platform to generate high-quality single-end reads of 101 nucleotides (some with 85 nucleotides due to machine failure) in length.

To survey the gene expression profile in the spleens of large yellow croakers induced by poly(I:C), a paired-end library was constructed according to the manufacturer's protocol. Polyadenylated RNA was isolated using the Oligotex mRNA Midi Kit (Qiagen Inc., Valencia, CA, USA). Two hundred nanograms of mRNA were used for the library preparation. The RNA-seq library was constructed using Illumina Whole Transcriptome Analysis Kit following the standard protocol (Illumina GA II Sequencing System). An 80 bp paired-end run was performed on the Illumina GAII platform (Illumina, Inc., San Diego, CA, USA) to assemble the whole transcriptome de novo. The generated sequence data have been submitted to the NCBI SRA database, and the accession number is SRP035897.