From 75336a20778fe27c22296843d277bdb32e861d33 Mon Sep 17 00:00:00 2001 From: jvfe Date: Mon, 2 Dec 2024 16:00:14 -0300 Subject: [PATCH] docs: Add analysis example --- docs/analysis_example.md | 94 ++++++++++++++++++++++ docs/assets/betadiv_microview.png | Bin 0 -> 38076 bytes docs/assets/fastp_filtered_reads_plot.png | Bin 0 -> 65287 bytes docs/assets/kaiju-topfive-plot.png | Bin 0 -> 80886 bytes mkdocs.yml | 1 + 5 files changed, 95 insertions(+) create mode 100644 docs/analysis_example.md create mode 100644 docs/assets/betadiv_microview.png create mode 100644 docs/assets/fastp_filtered_reads_plot.png create mode 100644 docs/assets/kaiju-topfive-plot.png diff --git a/docs/analysis_example.md b/docs/analysis_example.md new file mode 100644 index 0000000..947b3f2 --- /dev/null +++ b/docs/analysis_example.md @@ -0,0 +1,94 @@ +# Example analysis - Crohn's disease microbiome data + +To showcase the potential EURYALE has to expedite the analysis of microbiome data, let's +try analysing a publicly available gut microbiome dataset ([PRJNA175224](https://www.ncbi.nlm.nih.gov/bioproject/PRJNA175224)). +This dataset contains 7 gut microbiome samples from healthy donors and 4 from donors with Crohn's disease. + +Feel free to download the original dataset to a directory called `raw_data` and follow along. + +## Acquiring databases and running the pipeline + +Let's first download the databases and references. +For this analysis, we'll focus on the taxonomic classification results after de-contamination. +So, let's download the Human reference genome as well as Kaiju's database: + +```bash +nextflow run dalmolingroup/euryale \ + --download_kaiju \ + --download_host \ + --outdir references \ + -entry download \ + -profile singularity +``` + +Once that's done, let's execute the analysis on the data itself. +Your samplesheet should look something like this: + +```csv +sample,fastq_1,fastq_2 +SRR579274,raw_data/SRR579274_1.fastq.gz,raw_data/SRR579274_2.fastq.gz +SRR579275,raw_data/SRR579275_1.fastq.gz,raw_data/SRR579275_2.fastq.gz +SRR579276,raw_data/SRR579276_1.fastq.gz,raw_data/SRR579276_2.fastq.gz +SRR579277,raw_data/SRR579277_1.fastq.gz,raw_data/SRR579277_2.fastq.gz +SRR579278,raw_data/SRR579278_1.fastq.gz,raw_data/SRR579278_2.fastq.gz +SRR579279,raw_data/SRR579279_1.fastq.gz,raw_data/SRR579279_2.fastq.gz +SRR579280,raw_data/SRR579280_1.fastq.gz,raw_data/SRR579280_2.fastq.gz +SRR579281,raw_data/SRR579281_1.fastq.gz,raw_data/SRR579281_2.fastq.gz +SRR579290,raw_data/SRR579290_1.fastq.gz,raw_data/SRR579290_2.fastq.gz +SRR579291,raw_data/SRR579291_1.fastq.gz,raw_data/SRR579291_2.fastq.gz +SRR579292,raw_data/SRR579292_1.fastq.gz,raw_data/SRR579292_2.fastq.gz +``` + +And your command should look something like this one: + +```bash +nextflow run dalmolingroup/euryale \ + --input samplesheet.csv \ + --outdir results \ + --skip_alignment \ + --skip_functional \ + --kaiju_db references/kaiju_db_nr_2023-05-10.tgz \ + --host_fasta references/Homo_sapiens.GRCh38.dna.primary_assembly.fa.gz \ + -profile singularity \ + -r "v1.0.3" +``` + +Check the [parameter documentation](https://dalmolingroup.github.io/euryale/params/) for a full description +of possible parameters. + +- We're skipping the functional annotation section just to expedite the results, but feel free to include these + steps in your own analysis. + +Once that's all done, let's check the results we got. + +## Exploring the resulting data + +First, let's open the MultiQC report in our browser and take a look. +It should be available in `results/multiqc/multiqc_report.html`. + +We can first see that most reads (80%) in each sample passed +the quality filter, thankfully: + +![](assets/fastp_filtered_reads_plot.png) + +We can also see that most samples did not have their reads assigned +to any particular taxon: + +![](assets/kaiju-topfive-plot.png) + +This could be due to high host contamination, or could necessitate +the use of another database, or something else entirely! Either way, it's something worth investigating. +We can also see there is a strange spike of _Enterocloster boltae_ in one of the samples, which also +warrants further investigation. + +If we now check the MicroView results, available in `results/taxonomy/microview/microview_report.html`, +we can see further points of interest to investigate in our data. + +Let's see the Beta-diversity PCoA, for example: + +![](assets/betadiv_microview.png) + +The plot shows a somewhat strange division in the dataset: 4 samples stretch further in +the PC1 than the rest. This could be due to some biological variable or an error in the data generation process. +Either way, it's worth saving the PCoA table, available in `results/taxonomy/microview/microview_tables/` and +crossing the sample names with the metadata of this study. diff --git a/docs/assets/betadiv_microview.png b/docs/assets/betadiv_microview.png new file mode 100644 index 0000000000000000000000000000000000000000..e07c0ec3ce4099f28e7021247ec0337d2eb9754e GIT binary patch literal 38076 zcmeFacUV(f_b#d+CJKZgDgvQKMWrb<(lH3A2v`9T5J3THq4y?%h|(1hktTLTq)0Ck z6p*e+uaVxHbO>iI5Jh)6-~N8*xzD-x?!WdP%r&#-nrp5($9Tv44t>>B6lu3JZ{M_O z6YU8l`LmlgZAER`w3!Is3jVTlQD1V?CXP)fQgW1Upg+y z!L{|^BM*5rg_TkYVsZJV=8n{mP#5cp`GBLvc{BO1M@CcAybC5_x>z_W*&nrTc$)+jJs=smKR&GSh);abxqr{ls_$_e6K4exj8%9sIeVfUvor7;5+928I~ z-J~Ze1GSMdwNvqGb<=3n%h%QQlW>@p>z7PgVq9^(>4+chO`W1y3kT|kNkw};9OfML zC7qramtJf-atCVihnh?)>#H{Oy4b-)>Wo)pIEX~-zwz8@8y;IX9ihQSy7I{9Mz}2I zyjE1i2YSTym(T6!@z}R@5gI6{=_S;p9pl?W|Ff4re;{uBQ@rm^IE+E$eBKsf+~9&T 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literal 0 HcmV?d00001 diff --git a/mkdocs.yml b/mkdocs.yml index 48a02b0..f628e05 100644 --- a/mkdocs.yml +++ b/mkdocs.yml @@ -5,6 +5,7 @@ nav: - Home: README.md - Usage: usage.md - Output: output.md + - Analysis Example: analysis_example.md - Citations: CITATIONS.md - Reference: - Parameters: params.md