Nematostella High-density RNAseq time-course

Abstract

RNA-Seq was performed on Nematostella embryos at 20 timepoints: unfertilized eggs, 1hour post fertilization (hpf), 2hpf, 3hpf, 4hpf, 5hpf, 6hpf, 7hpf, 8hpf, 9hpf, 10hpf, 11hpf, 12hpf, 13hpf, 14hpf, 15hpf, 16hpf, 17hpf, 18hpf, 19hpf. Two samples (A and B) were collected for each time point. All embryos were fertilized at the same time and 300 embryos were collected for each sample. After harvesting the embroys, they were lysed and the mRNAs was extracted using Dynabeads (Invitrogen). Directional sequencing libraries were constructed using the ScriptSeq V2 kit (Epicentre). The libraries were size selected with PippinPrep to 300bp insert size. Two different control RNA sets from the National Institute of Standards and Technology (NIST) (ERCC Spike-in, Ambion) were added into the m-RNA prior to the library preparation. Libraries were sequenced using the Illumina HiSeq 1000, version 3 chemistry, 100bp paired-end reads and produced over 1.4 billion reads. The reads were quality controlled filtered and mapped against the Nematostella embryonic transcriptome (Tulin et al.). This database contains the raw read files for each library.