title: Illumina HiSeq 1000 sequencing; The Tetraodon Nigroviridis Transcriptome During Early Development.
design: The Tetraodon Nigroviridis Transcriptome During Early Development.
library_construction_protocol: Tetraodon embryos were obtained by in vitro fertilization of eggs and their development was observed through brightfield microscopy. Total RNA was extracted with Trizol (Invitrogen) according to the manufacturer s protocol from eggs, whole embryo at 30% epiboly (30 epi) and whole embryo at 24 hours post fertilisation (24 hpf). The RNA samples were treated with 2U Dnase I (Qiagen) per μg RNA sample at 37°C for 10 minutes. Digested samples were then treated with 20 mg/mL proteinase K (Sigma Aldrich) at 37°C for 45 minutes. The quality and quantity of total RNA were assessed with the Bioanalyzer 2100 (Agilent) and no sign of degradation was detected (RIN > 9.0). Sequencing libraries were generated from total RNA samples following the Truseq RNA protocol (Illumina).