Marine sponge Haliclona fulva hologenome sequences - total contigs assembled
<p>Dataset (multifasta) of DNA contig sequences assembled from Illumina metagenomic reads of Haliclona fulva hologenome. The sequences analyses are described in the peer-reviewed article at https://doi.org/10.3389/fmars.2021.736817 entitled: "The Hologenome of Haliclona fulva (Porifera, Demospongiae) Reveals an Abundant and Diverse Viral Community" by<br> Erika García-Bonilla1,2†, Diego Chaves-Moreno3†, Diego Riaño-Pachón4, Wilson Terán5, Alberto Acosta1 and Howard Junca2*<br> 1Laboratorio Ecosistemas Marinos Estratégicos, Unidad de Ecología y Sistemática - UNESIS, Facultad de Ciencias - Departamento de Biología, Pontificia Universidad Javeriana, Bogotá, Colombia<br> 2RG Microbial Ecology: Metabolism, Genomics & Evolution, Division of Ecogenomics & Holobionts, Microbiomas Foundation, Chía, Colombia<br> 3Microbial Interactions and Processes Research Group, Helmholtz Centre for Infection Research, Braunschweig, Germany<br> 4Laboratório de Biologia Computacional, Evolutiva e de Sistemas, Centro de Energía Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, Brazil<br> 5Biología de Plantas y Sistemas Productivos, Departamento de Biología, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia</p> <p>"Sample Collection</p> <p>Three specimens of <em>H. fulva</em> (HF1, HF2, and HF3) were collected by SCUBA diving in the Mediterranean Sea at the Grotte du Lido, in the bay of Villefranche-sur-Mer, France (latitude: 43° 41′ 31.49″ N; longitude: 7° 19′ 12.19″ E) at 35 m depth. After collection, all individual samples were placed in independent plastic bags. They were preserved in ethanol 70% (v/v) and stored at −20°C until further analysis. The samples were selected based on the findings of our previous study (<a href="https://www.frontiersin.org/articles/10.3389/fmars.2021.736817/full#B11">García-Bonilla et al., 2019</a>) and are coming from exactly the same spots and colonies/specimens analyzed regarding microbiome content by 16S amplicons, showing a stable and consistent microbial composition of low microbial abundance and diversity.</p> <p>DNA Extraction and Sequencing</p> <p>Metagenomic DNA was extracted from 15 g sponge wet weight using a MagAttract<sup>®</sup> HMW DNA kit (Qiagen, Germany) following the manufacturer’s instructions. Tissue lysis was made for 16 h according to protocol. Extracted DNA was eluted with 50 μl water, and its concentration determined by fluorescence using the Qubit<sup>®</sup> dsDNA BR assay kit (Thermo Fisher Scientific). Nucleic acid integrity was verified by 0.8% agarose gel electrophoresis. Sponge DNA extracts were used as template to perform isothermal multiple displacement hologenome amplification with a phi29 polymerase of high processivity and fidelity in order to increase total DNA concentrations (<a href="https://www.frontiersin.org/articles/10.3389/fmars.2021.736817/full#B36">Lasken, 2007</a>) and to avoid the effect of coextracted enzymatic inhibitors detected in <em>H. fulva</em>, improving further downstream processes (library construction and sequencing). The amplification process was made using REPLI-g Mini kit (Qiagen, Germany), according to manufacturer’s recommendations. Briefly, samples were incubated at 30°C for 11 h followed by 3 min at 65°C for polymerase inactivation. For all assays, a negative control was run to evaluate the presence of contaminants during amplification. Quantity and quality of DNA was measured as described previously.</p> <p>Genomic libraries were constructed using TruSeq DNA PCR free kit (Illumina, United States). Shotgun hologenome sequencing was performed using paired-end Illumina technology (Macrogen, South Korea). The complete raw sequencing data obtained is publicly accessible at NCBI GenBank SRA under Bioproject PRJNA741981.</p> <p>Raw Data Processing and Assembly</p> <p>An initial filtering of the sequencing reads was done after visual evaluation using FastQC (v.0.11.8) consisting of a quality and length-based filtering performed with Trimmomatic (v.0.31). Filtered and processed reads were assembled using Megahit (v.1.0) (<a href="https://www.frontiersin.org/articles/10.3389/fmars.2021.736817/full#B37">Li et al., 2014</a>) and IDBA-UD (<a href="https://www.frontiersin.org/articles/10.3389/fmars.2021.736817/full#B46">Peng et al., 2012</a>) with default parameters. To compare, identify and join overlaps between reads from each assembly, we used minimus2 from the AMOS suite with default parameters (<a href="https://www.frontiersin.org/articles/10.3389/fmars.2021.736817/full#B65">Treangen et al., 2011</a>). Obtained contigs were filtered and those <500 bp were removed."</p> <p> </p>
ShareScore
40/100
Overall dataset sharing score
Score breakdown
These five areas show where the dataset supports — or may limit — practical reuse.
- Stewardship
- 8
- Harmonization
- 4
- Access
- 16
- Reuse readiness
- 8
- Engagement
- 4