RNA-Seq of Wild Type Nematostella vectensis: Light Dark Conditions at 7am rep 2
Total RNA was extracted from all sampled polyps (n = 96) using TRIzol reagent (Invitrogen). Purified RNA samples were analyzed using a NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific) to assess RNA quantity and 2200 TapeStation (Agilent) to assess RNA quality (RNA integrity number, >8.5). From each of the 96 samples (4 biological replicates in LD and 3 in DD, with the highest-quality extracts  4 experimental subgroups  7 time points, 1.5 mg of RNA was sent for library preparation (INCPM mRNAseq protocol) and sequencing at the Weizmann institute, Israel. Libraries ran on nine lanes of an Illumina HiSeq2500 using the  multiplexing  strategy  of  the TruSeq protocol. The protocol starts with polyA selection that re-sults in mRNA selection only. On average, ~18 million paired-end reads, 100 bp long, were obtained for each sample. FastQC (www.bioinformatics.babraham.ac.uk/projects/fastqc/)  was  used  to  quality  check raw parsed fastq files before further analysis.