aanat2-deltaCLK pineal gland CT2
Overall, 14 libraries [12 time points from Tg(aanat2:EGFP-ΔCLK) fish and two control samples from Tg(aanat2:EGFP) fish] were run on a single flow cell of an Illumina HiSeq2500 machine (rapid run mode) using the multiplexing strategy of the TruSeq protocol (Institute of Applied Genomics, Italy).  Library construction was conducted using Illumina's TruSeq RNA Sample Prep Kit following the manufacture protocol. Briefly, starting from total RNA, the messenger RNA was purified using polyA selection, then chemically fragmented and converted into single-stranded cDNA using random hexamer priming. Afterwards, the second strand was generated to create double-stranded cDNA. Finally, adapters were added by ligation to the double-stranded cDNA, which prepared the samples for cluster generation. After library quality check by Bioanalyzer High Sensitivity chip, clusters were generated and libraries were run on HiSeq2500 by using standard Illumina sequencing workflow with the multiplexing option. Primary data analyses of sequence data and quality controls were performed using the Illumina pipeline . The reads were filtered using Illumina's HiSeq2500 softwares, Illumina indexes separation was also performed.