title: RNA-Seq of Stomolophus sp. 2: adult jellyfish bell tissue 18C_rep1
design: RNA samples were quantified using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked using Agilent  TapeStation 4200 (Agilent Technologies, Palo Alto, CA, USA). Sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina following manufacturers instructions (NEB, Ipswich, MA, USA). Briefly, mRNAs were first enriched with Oligo(dT) beads. Enriched mRNAs were fragmented for 15 minutes at 94 C. First strand and second strand cDNAs were subsequently synthesized. cDNA fragments were end-repaired and adenylated at 3ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by limited-cycle PCR. The sequencing libraries were validated on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified with Qubit 2.0, as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). The sequencing libraries (nine) were clustered on 1 lane of a flowcell. After clustering, the flowcell was loaded on the Illumina HiSeq 4000 instrument according to manufacturers instructions. The samples were sequenced using a 2x150bp Paired End (PE) configuration. Image analysis and base calling were conducted by the HiSeq Control Software (HCS). Raw sequence data (.bcl files) generated from Illumina HiSeq was converted into fastq files and de-multiplexed using Illumina's bcl2fastq 2.17 software. One mismatch was allowed for index sequence identification.