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topics/microbiome/tutorials/metagenomics-assembly/tutorial.md

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Metagenomics involves the extraction, sequencing and analysis of combined genomic DNA from **entire microbiome** samples. It includes then DNA from **many different organisms**, with different taxonomic background.
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Reconstructing the genomes of microorganisms in the sampled communities is critical step in analyzing metagenomic data. To do that, we can use **assembly** and assemblers, *i.e.* computational programs that stich together the small fragments of sequenced DNA produced by sequencing instruments.
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> <hands-on-title>Evaluation assembly quality with metaQUAST</hands-on-title>
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> 1. {% tool [QUAST](toolshed.g2.bx.psu.edu/repos/iuc/quast/quast/5.2.0+galaxy1) %} with parameters:
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> 1. {% tool [QUAST](toolshed.g2.bx.psu.edu/repos/iuc/QUAST/QUAST/5.2.0+galaxy1) %} with parameters:
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> - *"Assembly mode?*": `Individual assembly (1 contig file per samples)`
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> - *"Use customized names for the input files?"*: `No, use dataset names`
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> - {% icon param-collection %} *"Contigs/scaffolds file"*: output **MEGAHIT**
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> > 1. Import the metaQUAST report file from [Zenodo]({{ page.zenodo_link }}) or the Shared Data library:
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> >
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> > ```text
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> > {{ page.zenodo_link }}/files/QUAST_ERR2231567.html
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> > ```
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> >
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> {: .hands_on}
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> <details-title>Alternative ways to compute coverage</details-title>
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> 2 alternative ways to compute coverage are to
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> There are 2 alternative ways to compute coverage:
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> 1. Use [CoverM](https://github.com/wwood/CoverM), which is available in Galaxy
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> > <hands-on-title>Calculate coverage using CoverM</hands-on-title>
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QUAST identifies missassemblies by mapping the contigs to the reference genomes of the identified organisms. 3 types of misassemblies can be identified:
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![Image shows on the top a contig with a blue and a gren parts with white arrows (pointing on the right) on them and below a reference with 2 chromosomes. The 3 types of misassemblies are after schematized. Relocation: the blue and gren parts of the contig are on chr 1 but separated. Inversion: the blue and gren parts of the contig are on chr 1 but separated and with the arrows facing each other. Translocation: the blue part is on chr 1 and gren part on chr 2.](./images/quast_misassemblies.png "Source: <a href="https://quast.sourceforge.net/docs/manual.html#sec3.1.2">QUAST manual</a>"){:width="60%"}
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![Image shows on the top a contig with a blue and a gren parts with white arrows (pointing on the right) on them and below a reference with 2 chromosomes. The 3 types of misassemblies are after schematized. Relocation: the blue and gren parts of the contig are on chr 1 but separated. Inversion: the blue and gren parts of the contig are on chr 1 but separated and with the arrows facing each other. Translocation: the blue part is on chr 1 and gren part on chr 2.](./images/QUAST_misassemblies.png "Source: <a href="https://QUAST.sourceforge.net/docs/manual.html#sec3.1.2">QUAST manual</a>"){:width="60%"}
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1. **Relocation** occur based on signal from two mappings of the same contig against the same chromosome, with 2 cases:
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1. the 2 mappings are separated by an unmapped region of at least 1 kbp

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