Skip to content

Commit

Permalink
Merge pull request #2109 from shiltemann/update-abr
Browse files Browse the repository at this point in the history
[Metagenomics] Nanopore tutorial updates
  • Loading branch information
bgruening authored Oct 31, 2020
2 parents 705908a + b5e6ba3 commit c1ebf05
Show file tree
Hide file tree
Showing 5 changed files with 95 additions and 54 deletions.
1 change: 1 addition & 0 deletions _config.yml
Original file line number Diff line number Diff line change
Expand Up @@ -97,6 +97,7 @@ icon-tag:
feedback: far fa-comments
galaxy-barchart: fas fa-chart-bar
galaxy-chart-select-data: fas fa-database
galaxy-clear: fas fa-times-circle
galaxy-columns: fas fa-columns
galaxy-cross: fas fa-times
galaxy-dropdown: fas fa-caret-down
Expand Down
Binary file added shared/images/history_search_datasets.png
Loading
Sorry, something went wrong. Reload?
Sorry, we cannot display this file.
Sorry, this file is invalid so it cannot be displayed.
13 changes: 13 additions & 0 deletions snippets/history_search.md
Original file line number Diff line number Diff line change
@@ -0,0 +1,13 @@
>
> > ### {% icon tip %} Tip: Searching your history
> >
> > To make it easier to find datasets in large histories, you can filter your history by keywords as follows:
> > 1. **Click** on the *search datasets* box at the top of the history panel.
> >
> > ![history search box]({{site.baseurl}}/shared/images/history_search_datasets.png)
> >
> > 2. **Type** a search term in this box
> > - For example a tool name, or sample name
> > 3. To undo the filtering and show your full history again, press on the *clear search* button {% icon galaxy-clear %} next to the search box
> {: .tip}
>
16 changes: 16 additions & 0 deletions snippets/scratchbook.md
Original file line number Diff line number Diff line change
@@ -0,0 +1,16 @@
>
> > ### {% icon tip %} Tip: Using the Scratchbook to view multiple datasets at once
> > If you would like to view two or more datasets at once, you can use the **Scratchbook** feature in Galaxy:
> > 1. **Click** on the *Scratchbook* icon {% icon galaxy-scratchbook%} on the top menu bar.
> > - You should see a little checkmark on the icon now
> > 2. **View** {% icon galaxy-eye %} a dataset by clicking on the eye icon {% icon galaxy-eye %} to view the output
> > - You should see the output in a window overlayed over Galaxy
> > - You can resize this window by dragging the bottom-right corner
> > 3. **Click** outside the file to exit the Scratchbook
> > 4. **View** {% icon galaxy-eye %} a second dataset from your history
> > - You should now see a second window with the new dataset
> > - This makes it easier to compare the two outputs
> > 5. Repeat this for as many files as you would like to compare
> > 6. You can turn off the **Scratchbook** {% icon galaxy-scratchbook %} by clicking on the icon again
> {: .tip}
>
Original file line number Diff line number Diff line change
Expand Up @@ -128,10 +128,10 @@ report page.
> ### {% icon hands_on %} Hands-on: Plotting scripts for long read sequencing data
>
> 1. **NanoPlot** {% icon tool %} with the following parameters
> - *"Select multifile mode"*: `batch`
> - *"Type of the file(s) to work on"*: `fasta`
> - *"files"*: The `Plasmids` dataset collection you just created
> 1. {% tool [Nanoplot](tool toolshed.g2.bx.psu.edu/repos/iuc/nanoplot/nanoplot/1.28.2+galaxy1) %} with the following parameters
> - {% icon param-select %} *"Select multifile mode"*: `batch`
> - {% icon param-select %} *"Type of the file(s) to work on"*: `fasta`
> - {% icon param-collection %} *"files"*: The `Plasmids` dataset collection you just created
>
> {% include snippets/select_collection.md %}
>
Expand Down Expand Up @@ -178,14 +178,14 @@ the Minimap2 publication ({% cite Li2018 %}).
> ### {% icon hands_on %} Hands-on: Pairwise sequence alignment
>
> 1. **Map with minimap2** {% icon tool %} with the following parameters
> - *"Will you select a reference genome from your history or use a built-in index?"*: `Use a genome from history and build index`
> - *"Use the following data collection as the reference sequence"*: `Created dataset collection (Plasmids)`
> - *"Single or Paired-end reads"*: `Single`
> - *"Select fastq dataset"*: The `Plasmids` dataset collection
> - *"Select analysis mode (sets default)"*: `Oxford Nanopore all-vs--all overlap mapping`
> 1. {% tool [Map with minimap2](toolshed.g2.bx.psu.edu/repos/iuc/minimap2/minimap2/2.17+galaxy2) %} with the following parameters
> - {% icon param-select %} *"Will you select a reference genome from your history or use a built-in index?"*: `Use a genome from history and build index`
> - {% icon param-collection %} *"Use the following data collection as the reference sequence"*: `Plasmids` dataset collection we just created
> - {% icon param-select %} *"Single or Paired-end reads"*: `Single`
> - {% icon param-collection %} *"Select fastq dataset"*: The `Plasmids` dataset collection
> - {% icon param-select%} *"Select a profile of preset options"*: `Oxford Nanopore all-vs--all overlap mapping`
> - In the section **Set advanced output options**:
> - *"Select an output format"*: `paf`
> - {% icon param-select %} *"Select an output format"*: `paf`
>
> {% include snippets/select_collection.md %}
>
Expand Down Expand Up @@ -231,9 +231,9 @@ Thus the per-base error rate is similar to the raw input reads.
> ### {% icon hands_on %} Hands-on: De novo assembly
>
> 1. **miniasm** {% icon tool %} with the following parameters
> - *"Sequence Reads"*: The `Plasmids` dataset collection
> - *"PAF file"*: `Output Minimap dataset collection` created by **Minimap2** {% icon tool %}
> 1. {% tool [miniasm](toolshed.g2.bx.psu.edu/repos/iuc/miniasm/miniasm/0.3+galaxy0) %} with the following parameters
> - {% icon param-collection %} *"Sequence Reads"*: The `Plasmids` dataset collection
> - {% icon param-collection %} *"PAF file"*: `Output Minimap dataset collection` created by **Minimap2** {% icon tool %}
>
> {% include snippets/select_collection.md %}
>
Expand All @@ -256,33 +256,34 @@ Remapping is done with the original reads, using the Miniasm assembly as a refer
> ### {% icon hands_on %} Hands-on: Pairwise sequence alignment
>
> 1. **GFA to Fasta** {% icon tool %} with the following parameters
> - *"Input GFA file"*: the `Assembly Graph` (collection) created by **Miniasm** {% icon tool %}
>
> 2. **Map with minimap2** {% icon tool %} with the following parameters
> - *"Will you select a reference genome from your history or use a built-in index?"*: `Use a genome from history and build index`
> - *"Use the following dataset as the reference sequence"*: `FASTA file` collection created by **GFA to Fasta** {% icon tool %}
> - *"Single or Paired-end reads"*: `single`
> - *"Select fastq dataset"*: The `Plasmids` collection
> - *"Select analysis mode (sets default)"*: `PacBio/Oxford Nanopore read to reference mapping (-Hk19)`
> - In the section **Set advanced output options**:
> - *"Select an output format"*: `paf`
>
>
> {% include snippets/select_collection.md %}
> 1. {% tool [GFA to Fasta](toolshed.g2.bx.psu.edu/repos/iuc/gfa_to_fa/gfa_to_fa/0.1.1) %} with the following parameters
> - {% icon param-collection %} *"Input GFA file"*: the `Assembly Graph` (collection) created by **Miniasm** {% icon tool %}
>
> > ### {% icon question %} Question
> >
> > How many contigs do we have for the RB05 sample after the use of **Minimap2** {% icon tool %} and **Miniasm** {% icon tool %}?
> > How many contigs do we have for the RB05 sample after de novo assembly?
> > <br><br>
> > Hint: run **Nanoplot** {% icon tool %} on the output of **GFA to Fasta** {% icon tool %}
> >
> > > ### {% icon solution %} Solution
> > > 22
> > > 25
> > >
> > > This can be determined by looking at the NanoStats output of NanoPlot.
> > {: .solution }
> {: .question}
>
> 2. {% tool [Map with minimap2](toolshed.g2.bx.psu.edu/repos/iuc/minimap2/minimap2/2.17+galaxy2) %} with the following parameters
> - {% icon param-select %} *"Will you select a reference genome from your history or use a built-in index?"*: `Use a genome from history and build index`
> - {% icon param-collection %} *"Use the following dataset as the reference sequence"*: `FASTA file` output from **GFA to Fasta** {% icon tool %} (collection)
> - {% icon param-select %} *"Single or Paired-end reads"*: `single`
> - {% icon param-collection %} *"Select fastq dataset"*: The `Plasmids` collection
> - {% icon param-select %} *"Select a profile of preset options"*:: `PacBio/Oxford Nanopore read to reference mapping (-Hk19)`
> - In the section **Set advanced output options**:
> - {% icon param-select %} *"Select an output format"*: `paf`
>
>
> {% include snippets/select_collection.md %}
>
{: .hands_on}
## Ultrafast consensus module using Racon
Expand All @@ -296,10 +297,10 @@ It supports data produced by both Pacific Biosciences and Oxford Nanopore Techno
> ### {% icon hands_on %} Hands-on: Consensus module
>
> 1. **Racon** {% icon tool %} with the following parameters
> - *"Sequences"*: The `Plasmids` dataset collection
> - *"Overlaps"*: the latest `PAF file` collection created by **Minimap2** {% icon tool %}
> - *"Target sequences"*: the `FASTA file` collection created by **GFA to Fasta** {% icon tool %}
> 1. {% tool [Racon](toolshed.g2.bx.psu.edu/repos/bgruening/racon/racon/1.4.13) %} with the following parameters
> - {% icon param-collection %} *"Sequences"*: The `Plasmids` dataset collection
> - {% icon param-collection %} *"Overlaps"*: the latest `PAF file` collection created by **Minimap2** {% icon tool %}
> - {% icon param-collection %} *"Target sequences"*: the `FASTA file` collection created by **GFA to Fasta** {% icon tool %}
>
{: .hands_on}
Expand All @@ -326,8 +327,8 @@ By visualizing these assembly graphs, Bandage allows users to better understand,
> ### {% icon hands_on %} Hands-on: Visualising de novo assembly graphs
>
> 1. **Bandage image** {% icon tool %} with the following parameters
> - *"Graphical Fragment Assembly"*: the `Assembly graph` collection created by **Miniasm** {% icon tool %}
> 1. {% tool [Bandage image](toolshed.g2.bx.psu.edu/repos/iuc/bandage/bandage_image/0.8.1+galaxy2) %} with the following parameters
> - {% icon param-collection %} *"Graphical Fragment Assembly"*: the `Assembly graph` collection created by **Miniasm** {% icon tool %}
>
> 2. Explore {% icon galaxy-eye %} the output images
>
Expand Down Expand Up @@ -372,23 +373,34 @@ Let's try it on our data!
> ### {% icon hands_on %} Hands-on: Unicycler assembly
>
> 1. **Create assemblies with Unicycler** {% icon tool %} with the following parameters
> - *"Paired or Single end data"*: `None`
> - *"Select long reads. If there are no long reads, leave this empty"*: The `Plasmids` dataset collection
> 1. {% tool [Create assemblies with Unicycler](toolshed.g2.bx.psu.edu/repos/iuc/unicycler/unicycler/0.4.8.0) %} with the following parameters
> - {% icon param-select %} *"Paired or Single end data"*: `None`
> - {% icon param-collection %} *"Select long reads. If there are no long reads, leave this empty"*: The `Plasmids` dataset collection
>
> 2. **Bandage image** {% icon tool %} with the following parameters
> - *"Graphical Fragment Assembly"*: the `Final Assembly Graph` collection created by **Unicycler** {% icon tool %}
> 2. {% tool [Bandage image](toolshed.g2.bx.psu.edu/repos/iuc/bandage/bandage_image/0.8.1+galaxy2) %} with the following parameters
> - {% icon param-collection %} *"Graphical Fragment Assembly"*: the `Final Assembly Graph` collection created by **Unicycler** {% icon tool %}
>
> 3. Examine {% icon galaxy-eye %} the output images again
>
> > ### {% icon question %} Question
> >
> > For which samples has the plasmid assembly improved?
> >
> > > ### {% icon solution %} Solution
> > > Exploring the outputs for all the samples reveals that many now display circular assemblies, indicating the full plasmids sequence was resolved.
> > {: .solution }
> {: .question}
> 4. Use the **Scratchbook** {% icon galaxy-scratchbook %} to compare the two assemblies for sample `RB01`
> - Compare the **Bandage** {% icon tool %} images for our two assemblies:
> 1. The assembly we got from running **minimap2, miniasm, racon** {% icon tool%} (first time we ran bandage)
> 2. The assembly obtained with **Unicycler** {% icon tool %}
> - Tip: Search your history for the term `bandage` to easily find the outputs from our two bandage runs
>
> {% include snippets/scratchbook.md %}
> {% include snippets/history_search.md %}
>
> 5. Repeat this comparison for the other samples.
>
> > ### {% icon question %} Question
> >
> > For which samples has the plasmid assembly improved?
> >
> > > ### {% icon solution %} Solution
> > > Exploring the outputs for all the samples reveals that many now display circular assemblies, indicating the full plasmids sequence was resolved.
> > {: .solution }
> {: .question}
>
{: .hands_on}
Expand All @@ -410,8 +422,8 @@ It relies on the neural network models trained on full genome and plasmid sequen
> ### {% icon hands_on %} Hands-on: Prediction of plasmid sequences
>
> 1. **PlasFlow** {% icon tool %} with the following parameters
> - *"Sequence Reads"*: the `Final Assembly` collection created by **Unicycler** {% icon tool %}
> 1. {% tool [PlasFlow](toolshed.g2.bx.psu.edu/repos/iuc/plasflow/PlasFlow/1.0) %} with the following parameters
> - {% icon param-collection %} *"Sequence Reads"*: the `Final Assembly` collection created by **Unicycler** {% icon tool %}
>
> > ### {% icon question %} Question
> >
Expand Down Expand Up @@ -456,8 +468,8 @@ and compiles a summary report of detected antimicrobial resistance genes.
> ### {% icon hands_on %} Hands-on: Prediction of AMR genes
>
> 1. **staramr** {% icon tool %} with the following parameters
> - *"genomes"*: the `Final Assembly` collection created by **Unicycler**
> 1. {% tool [staramr](toolshed.g2.bx.psu.edu/repos/nml/staramr/staramr_search/0.7.1+galaxy2) %} with the following parameters
> - {% icon param-collection %} *"genomes"*: the `Final Assembly` collection created by **Unicycler** {% icon tool %}
>
> > ### {% icon question %} Question
> >
Expand Down Expand Up @@ -518,4 +530,3 @@ You have worked your way through the following pipeline:
![Workflow representation of this tutorial](../../images/plasmid-metagenomics-nanopore/Workflow.png)

0 comments on commit c1ebf05

Please sign in to comment.