Bell Center Data Sets

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https://hdl.handle.net/1912/5591

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Now showing 1 - 4 of 4
  • Dataset
    GRIPseq data for S. Purpuratus 24 hpf
    ( 2015-01-19) Tulin, Sarah ; Barsi, Julius C. ; Bocconcelli, Carlo ; Smith, Joel
    This dataset is the result of a new method for identifying putative cis-regulatory elements genome wide, termed GRIPseq for Genome-wide Regulatory element Immunoprecipitation. Our protocol combines elements of chromatin conformation capture (e.g., 3C, 4C, Hi-C), chromatin immunoprecipitation, and paired-end high-throughput sequencing with molecular tools that enrich for active cis-regulatory elements across the genome. Our pioneer dataset, available to the community, derives from the purple sea urchin, Stronglyocentrotus purpuratus, mesenchyme blastula embryos collected at 24 hpf. We benchmark our results against independent findings, i.e., rigorously tested cis-elements previously identified and find good congruence at the loci we have investigated, and in particular we find a strong signal-to-noise ratio for known distal elements, marking an improvement over existing methods. A corollary of this approach is the ability, in many cases, to link cis-regulatory elements to the genes they regulate.
  • Dataset
    Nematostella vectensis BAC sequences
    ( 2013-07-11) Fischer, Antje H. L. ; Tulin, Sarah ; Fredman, David ; Smith, Joel
    A key tool for investigating the regulation of genes is represented by Bacterial Artificial Chromosomes-reporter constructs (BAC). BACs are large insert libraries, often >>100 kb, which thus capture the genomic sequences surrounding a gene of interest, including all, or nearly all, of the elements underpinning regulation. Recombinant BACs, containing a reporter gene in place of the endogenous coding sequence of genes, can be utilized to drive the expression of reporter genes under the regulatory control of the gene of interest while still embedded within its genomic context. Systematic deletions within the BAC reporter construct can be used to identify the minimal reporter in an unbiased way, avoiding the risk of overlooking regulatory elements that may be many kilobases away from the transcription start-site. Nematostella vectensis (Edwardsiidae, Anthozoa, Cnidaria) has become an important model in regenerative biology, ecology, and especially in studies of evo-devo and gene-regulatory networks due to its interesting phylogenetic position and amenability to molecular techniques. This dataset represents the sequencing of 5 Nematostella vectensis BACs containing the genes of interest: NvDpp, NvChordin, NvFoxa, NvWnta, and NvWnt1.
  • Dataset
    Nematostella High-density RNAseq time-course
    ( 2013-06-14) Fischer, Antje H. L. ; Smith, Joel
    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.
  • Dataset
    Nematostella Embryonic Transcriptome
    ( 2012-12-12) Tulin, Sarah ; Aguiar, Derek ; Istrail, Sorin ; Smith, Joel
    RNA-Seq was performed on Nematostella Embryos at 5 timepoints during early development: 0hrs, 6hrs, 12hrs, 18hrs, 24hrs after fertilization. Embryos were harvested, lysed and mRNAs were selected using Dynabeads. Directional sequencing libraries were constructed using the ScriptSeq kit from Epicentre. A control RNA set from the National Institute of Standards and Technology (NIST) were spiked-into the library preparation. Libraries were sequenced using the Illumina HiSeq 1000, version 3 chemistry, 100bp paired-end reads and produced over 200million reads. The reads were quality controlled filtered and assembled using the Trinity Assembler. This database contains the raw read files and the assembled Transcriptome.