Overview

• This is a short introduction to the Galaxy user interface - the web page that you interact with.
• We will cover key tasks in Galaxy: uploading files, using tools, viewing histories, and running workflows.

Agenda

1. Overview
   1. What does Galaxy look like?
2. Key Galaxy actions
   1. Name your current history
   2. Upload a file
   3. Use a tool
   4. View results
   5. Run another tool
   6. Re-run that tool with changed settings
   7. Share your history
   8. Convert your analysis history into a workflow
   9. Create a new history
   10. Look at all your histories
   11. Run workflow in the new history
3. Conclusion

What does Galaxy look like?

Hands-on: Log in to Galaxy

1. Open your favorite browser (Chrome, Safari or Firefox as your browser, not Internet Explorer!)
2. Browse to your Galaxy instance
3. Log in or register

Comment: Different Galaxy servers

This is an image of Galaxy Australia, located at usegalaxy.org.au

The particular Galaxy server that you are using may look slightly different and have a different web address:

• The main Galaxy server is usegalaxy.org
• The European Galaxy server is usegalaxy.eu

You can also find more possible Galaxy servers at the top of this tutorial in Available on these Galaxies

The Galaxy homepage is divided into three panels:

• Tools on the left
• Viewing panel in the middle
• History of analysis and files on the right

The first time you use Galaxy, there will be no files in your history panel.

Key Galaxy actions

Name your current history

Your “History” is in the panel at the right.

Hands-on: Name history

1. Go to the History panel (on the right)

2. Click on galaxy-pencil (Edit) next to the history name (which by default is “Unnamed history”)

   

   Comment

   In some previous versions of Galaxy, you will need to click on the history name to rename it as shown here:

3. Type in a new name, for example, “My Analysis”
4. Click on Save

Comment: Renaming not an option?

If renaming does not work, it is possible you aren’t logged in, so try logging in to Galaxy first. Anonymous users are only permitted to have one history, and they cannot rename it.

Upload a file

Your “Tools” are in the panel at the left.

Hands-on: Upload a file from URL

1. At the top of the Tools panel (on the left), click galaxy-upload Upload

   

   This brings up a box:

   

2. Click Paste/Fetch data

3. Paste in the address of a file:

   https://zenodo.org/record/582600/files/mutant_R1.fastq
   

4. Click Start
5. Click Close

Your uploaded file is now in your current history. When the file has uploaded to Galaxy, it will turn green.

Comment

After this you will see your first history item (called a “dataset”) in Galaxy’s right panel. It will go through the gray (preparing/queued) and yellow (running) states to become green (success).

Details: Is this step taking a while?

Sometimes during courses, data upload gets a little slow. You can also import data through a history link.

1. Import history from: example input history

   Tip: Importing a history

   1. Open the link to the shared history
   2. Click on the new-history Import history button on the top right
   3. Enter a title for the new history
   4. Click on Import

2. Rename galaxy-pencil the the history to your name of choice.

What is this file?

Hands-on: View the dataset content

1. Click on the galaxy-eye (eye) icon next to the dataset name, to look at the file content

   

The contents of the file will be displayed in the central Galaxy panel.

This file contains DNA sequencing reads from a bacteria, in FASTQ format:

Open image in new tab

Use a tool

Let’s look at the quality of the reads in this file.

Hands-on: Use a tool

1. Type FastQC in the tools panel search box (top)

2. Click on the FastQC ( Galaxy version 0.73+galaxy0) tool

   The tool will be displayed in the central Galaxy panel.

3. Select the following parameters:
   • param-file “Raw read data from your current history”: the FASTQ dataset that we uploaded
   • No change in the other parameters
4. Click Execute

This tool will run and two new output datasets will appear at the top of your history panel.

View results

We will now look at the output dataset called FastQC on data 1: Webpage.

Comment

• Note that Galaxy has given this dataset a name according to both the tool name (“FastQC”) and the input (“data 1”) that it used.
• The name “data 1” means the dataset number 1 in Galaxy’s current history (our FASTQ file).

Hands-on: View results

• Once it’s green, click on the galaxy-eye (eye) icon next to the “Webpage” output dataset.

  The information is displayed in the central panel

  

This tool has summarised information about all of the reads in our FASTQ file.

Question

1. What was the length of the reads in the input FASTQ file?
2. Do these reads have higher quality scores in the centre or at the ends?

Solution

1. 150 bp
2. In the center

Run another tool

Let’s run a tool to filter out lower-quality reads from our FASTQ file.

Hands-on: Run another tool

1. Type Filter by quality in the tools panel search box (top)
2. Click on the tool Filter by quality ( Galaxy version 1.0.2+galaxy0)
3. Set the following parameters:
   • param-file “Input FASTQ file”: our initial FASTQ dataset
   • “Quality cut-off value”: 35
   • “Percent of bases in sequence that must have quality equal to / higher than cut-off value”: 80
4. Click Execute

After the tool has run, its output dataset will appear at the top of your History panel.

• This dataset will be called “Filter by quality on data 1”.
• Remember that Galaxy has named this file according to the tool it used (“Filter by quality”) and the input dataset (“data 1”).
• The actual numbers in front of the datasets in the history are not important.

What are the results from this filtering tool?

We could click on the eye icon to view the contents of this output file, but it will not be very informative - we will just see a list of reads.

Hands-on: Get metadata about a file

1. Click on the output dataset name in the History panel.

   This expands the information about the file.

   

Question

How many read has been discarded

Solution

1786 low-quality reads were discarded

Re-run that tool with changed settings

We can now try to filter our input reads to an even higher standard, and see how this changes the resulting output (an exploratory analysis). We will change the filter settings and re-run the tool.

Hands-on: Re-run the tool

1. Click on the galaxy-refresh icon (Run this job again) for the output dataset of Filter by quality tool

   

   This brings up the tool interface in the central panel with the parameters set to the values used previously to generate this dataset.

2. Change the settings to something even stricter

   For example, you might decide you want 80 percent of bases to have a quality of 36 or higher, instead of 35.

3. Click Execute

4. View the results: Click on the output dataset name to expand the information

   Comment

   Not the galaxy-eye (eye) icon.

Question

How many reads were discarded under these new filtering conditions?

Solution

If you selected 80% of bases with 36 as quality cut-off, then 11517 reads (92%) should have been discarded, which indicates that we have gone too far with the filtering in this case.

You can re-run a tool many times with different settings. Each time you re-run the tool, its new output datasets will appear at the top of your current history.

Share your history

Finally, let’s imagine that you had a problem in your analysis and you want to ask for help. The easiest way to ask for help is to share your history. Try and create a link for your history and share it with…yourself!

Tip: Sharing your History

Sharing your history allows others to import and access the datasets, parameters, and steps of your history.

1. Share via link
   • Open the History Options galaxy-gear menu (gear icon) at the top of your history panel and select Share or Publish
     • galaxy-toggle Make History accessible
     • A Share Link will appear that you give to others
   • Anybody who has this link can view and copy your history
2. Publish your history
   • galaxy-toggle Make History publicly available in Published Histories
   • Anybody on this Galaxy server will see your history listed under the Shared Data menu
3. Share only with another user.
   • Click the Share with a user button at the bottom
   • Enter an email address for the user you want to share with
   • Your history will be shared only with this user.
4. Finding histories others have shared with me
   • Click on User menu on the top bar
   • Select Histories shared with me
   • Here you will see all the histories others have shared with you directly

Note: If you want to make changes to your history without affecting the shared version, make a copy by going to galaxy-gear History options icon in your history and clicking Copy

Convert your analysis history into a workflow

When you look carefully at your history, you can see that it contains all the steps of our analysis, from the beginning (at the bottom) to the end (on top). The history in Galaxy records details of every tool you run and preserves all parameter settings applied at each step. But when you need to analyze new data, it would be tedious to do each step one-by-one again. Wouldn’t it be nice to just convert this history into a workflow that we will be able to execute again and again?

Galaxy makes this very easy with the Extract workflow option. This means any time you want to build a workflow, you can just perform the steps once manually, and then convert it to a workflow, so that next time it will be a lot less work to do the same analysis.

Hands-on: Extract workflow

1. Clean up your history: remove any failed (red) jobs from your history by clicking on the galaxy-delete button.

   This will make the creation of the workflow easier.

2. Click on galaxy-history-options (History options) at the top of your history panel and select Extract workflow.

   

   The central panel will show the content of the history in reverse order (oldest on top), and you will be able to choose which steps to include in the workflow.

   

3. Replace the Workflow name to something more descriptive, for example: QC and filtering.

4. Rename the workflow input in the box at the top of second column to: FASTQ reads

5. If there are any steps that shouldn’t be included in the workflow, you can uncheck them in the first column of boxes. In this case, uncheck the second Filter by quality tool at the bottom, where we used a too high quality cut-off.

6. Click on the Create Workflow button near the top.

   You will get a message that the workflow was created.

In a minute we will see how to find the extracted workflow and how to use it.

Create a new history

Let’s create a new history.

Hands-on: New history

1. Create a new history

   Tip: Creating a new history

   Click the new-history icon at the top of the history panel.

   If the new-history is missing:

   1. Click on the galaxy-gear icon (History options) on the top of the history panel
   2. Select the option Create New from the menu

2. Rename your history, e.g. “Next Analysis”

   Tip: Renaming a history

   1. Click on galaxy-pencil (Edit) next to the history name (which by default is “Unnamed history”)
   2. Type the new name
   3. Click on Save

   If you do not have the galaxy-pencil (Edit) next to the history name:

   1. Click on Unnamed history (or the current name of the history) (Click to rename history) at the top of your history panel
   2. Type the new name
   3. Press Enter

This new history does not have any datasets in it yet.

Look at all your histories

Where is your first history, called “My Analysis”?

Hands-on: View histories

1. Click on galaxy-history-options (History options) and then click on the galaxy-columns Show Histories side-by-side

   

   A new page will appear with all your histories displayed here.

2. Copy a dataset into your new history
   1. Click on the FASTQ dataset in “My Analysis” history
   2. Drag it into the “Next Analysis” history

   Open image in new tab
   
   

   This makes a copy of the dataset in the new history (without actually using additional disk space).

3. Click on the Home icon galaxy-home (or Analyze Data on older versions of Galaxy) in the top panel to go back to your analysis window

Your main Galaxy window will now show “Next Analysis” as the current history, and it will have one dataset in it.

At any time, you can go back into the “View all histories” page and “Switch to” a different history.

Run workflow in the new history

Now that we have built our workflow, let’s use it to re-create our small analysis in a single step. The same workflow could also be used on some new FASTQ data to quickly repeat the same analysis on different inputs.

Hands-on: Run workflow

1. Click on Workflow in the top menu bar of Galaxy.

   Here you have a list of all your workflows. Your newly created workflow should be listed at the top:

   

   If you click on a workflow name, you can see all available actions for the workflow, e.g. edit, copy, rename, delete.

2. Click on the workflow-run (Run workflow) button next to your workflow.

   The central panel will change to allow you to configure and launch the workflow.

   

3. Check that the “FASTQ reads” input is set to the FASTQ dataset we have copied to the new history.

   In this page we could change any parameter for the tools composing the workflow as we would do when running them one by one.

4. Click the Run Workflow button at the top-right of the screen.

   You should see a message that the workflow was successfully invoked. Then jobs will start to run and datasets appear in your “Next Analysis” history, replicating the steps of your previous history.

Conclusion

Well done! You have completed the short introduction to Galaxy, where you named the history, uploaded a file, used a tool, viewed results and run a workflow. Additional tutorials are available for a more in-depth introduction to Galaxy’s features.

Key points

• The Galaxy interface has tools on the left, viewing pane in the middle, and a history of your data analysis on the right.

• You can create a new history for each analysis. All your histories are saved.

• To get data into Galaxy, you can upload a file by pasting in a web address. There are other ways to get data into Galaxy (not covered in this tutorial): you can upload a file from your computer, and you can import an entire history.

• Choose a tool and change any settings for your analysis.

• Run the tool. The output files will be saved at the top of your history.

• View the output files by clicking on the eye icon.

• View all your histories and move files between them. Switch to a different history.

• Log out of your Galaxy server. When you log back in (to the same server), your histories will all be there.

Frequently Asked Questions

Feedback

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Citing this Tutorial

1. Anna Syme, Nicola Soranzo, A short introduction to Galaxy (Galaxy Training Materials). https://training.galaxyproject.org/training-material/topics/introduction/tutorials/galaxy-intro-short/tutorial.html Online; accessed TODAY
2. Hiltemann, Saskia, Rasche, Helena et al., 2023 Galaxy Training: A Powerful Framework for Teaching! PLOS Computational Biology 10.1371/journal.pcbi.1010752
3. Batut et al., 2018 Community-Driven Data Analysis Training for Biology Cell Systems 10.1016/j.cels.2018.05.012

BibTeX

@misc{introduction-galaxy-intro-short,
author = "Anna Syme and Nicola Soranzo",
	title = "A short introduction to Galaxy (Galaxy Training Materials)",
	year = "",
	month = "",
	day = ""
	url = "\url{https://training.galaxyproject.org/training-material/topics/introduction/tutorials/galaxy-intro-short/tutorial.html}",
	note = "[Online; accessed TODAY]"
}
@article{Hiltemann_2023,
	doi = {10.1371/journal.pcbi.1010752},
	url = {https://doi.org/10.1371%2Fjournal.pcbi.1010752},
	year = 2023,
	month = {jan},
	publisher = {Public Library of Science ({PLoS})},
	volume = {19},
	number = {1},
	pages = {e1010752},
	author = {Saskia Hiltemann and Helena Rasche and Simon Gladman and Hans-Rudolf Hotz and Delphine Larivi{\`{e}}re and Daniel Blankenberg and Pratik D. Jagtap and Thomas Wollmann and Anthony Bretaudeau and Nadia Gou{\'{e}} and Timothy J. Griffin and Coline Royaux and Yvan Le Bras and Subina Mehta and Anna Syme and Frederik Coppens and Bert Droesbeke and Nicola Soranzo and Wendi Bacon and Fotis Psomopoulos and Crist{\'{o}}bal Gallardo-Alba and John Davis and Melanie Christine Föll and Matthias Fahrner and Maria A. Doyle and Beatriz Serrano-Solano and Anne Claire Fouilloux and Peter van Heusden and Wolfgang Maier and Dave Clements and Florian Heyl and Björn Grüning and B{\'{e}}r{\'{e}}nice Batut and},
	editor = {Francis Ouellette},
	title = {Galaxy Training: A powerful framework for teaching!},
	journal = {PLoS Comput Biol} Computational Biology}
}

                   

Galaxy Administrators: Install the missing tools

You can use Ephemeris's shed-tools install command to install the tools used in this tutorial.

shed-tools install [-g GALAXY] [-a API_KEY] -t <(curl https://training.galaxyproject.org/training-material/api/topics/introduction/tutorials/galaxy-intro-short/tutorial.json | jq .admin_install_yaml -r)

Alternatively you can copy and paste the following YAML

---
install_tool_dependencies: true
install_repository_dependencies: true
install_resolver_dependencies: true
tools:
- name: fastq_quality_filter
  owner: devteam
  revisions: 43a7370aa010
  tool_panel_section_label: FASTA/FASTQ
  tool_shed_url: https://toolshed.g2.bx.psu.edu/
- name: fastq_quality_filter
  owner: devteam
  revisions: ce9cd02d5b88
  tool_panel_section_label: FASTA/FASTQ
  tool_shed_url: https://toolshed.g2.bx.psu.edu/
- name: fastqc
  owner: devteam
  revisions: 9da02be9c6cc
  tool_panel_section_label: FASTA/FASTQ
  tool_shed_url: https://toolshed.g2.bx.psu.edu/
- name: fastqc
  owner: devteam
  revisions: 3d0c7bdf12f5
  tool_panel_section_label: FASTA/FASTQ
  tool_shed_url: https://toolshed.g2.bx.psu.edu/