How Snakemake plans its jobs

Overview

Questions

• How do I visualise a Snakemake workflow?
• How does Snakemake avoid unecessary work?
• How do I control what steps will be run?

Objectives

• View the DAG for our pipeline
• Understand the logic Snakemake uses when running and re-running jobs

For reference, this is the Snakefile you should have to start the episode.

The DAG

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You may have noticed that one of the messages Snakemake always prints is:

OUTPUT

Building DAG of jobs...

A DAG is a Directed Acyclic Graph and it can be pictured like so:

The above DAG is based on four of our existing rules, and shows all the jobs Snakemake would run to trim, count and quantify the ref1 sample.

Note that:

• A rule can appear more than once, with different wildcards (a rule plus wildcard values defines a job)
• A rule may not be used at all, if it is not required for the target outputs
• The arrows show dependency ordering between jobs
• Snakemake can run the jobs in any order that doesn’t break dependency - for example kallisto_quant cannot run until both kallisto_index and trimreads have completed, but it may run before or after countreads
• This is a work list, not a flowchart, so there are no if/else decisions or loops - Snakemake runs every job in the DAG exactly once
• The DAG depends both on the Snakefile and on the requested target outputs, and the files already present
• When building the DAG, Snakemake does not look at the shell part of the rules at all - only when running the DAG will Snakemake check that the shell commands are working and producing the expected output files

How many jobs?

If we asked Snakemake to run kallisto_quant on all three of the reference samples (ref1, ref2, ref3), how many jobs would that be in total?

Show me the solution

10 in total: 3 * kallisto_quant // 6 * trimreads // 1 * kallisto_index // 0 * countreads

Snakemake is lazy, and laziness is good

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For the last few episodes, we’ve told you to run Snakemake like this:

BASH

$ snakemake -j1 -F -p desired_output_file

As a reminder, the -j1 flag tells Snakemake to run one job at a time, and -p is to print out the shell commands before running them.

The -F flag turns on forceall mode, and in normal usage you don’t want this.

At the end of the last chapter, we generated some kallisto results by running:

BASH

$ snakemake -j1 -F -p kallisto.temp33_1/abundance.h5

Now try without the -F option. Assuming that the output files are already created, you’ll see this:

BASH

$ snakemake -j1 -p kallisto.temp33_1/abundance.h5
Building DAG of jobs...
Nothing to be done.
Complete log: /home/zenmaster/data/yeast/.snakemake/log/2021-04-23T172441.519500.snakemake.log

In normal operation, Snakemake only runs a job if:

1. A target file you explicitly requested to make is missing
2. An intermediate file is missing and it is needed in the process of making a target file
3. Snakemake can see an input file which is newer than an output file
4. A rule definition or configuration has changed since the output file was created

The last of these relies on a ledger that Snakemake saves into the .snakemake directory.

Let’s demonstrate each of these in turn, by altering some files and re-running Snakemake without the -F option.

BASH

$ rm -rv kallisto.temp33_1
$ snakemake -j1 -p kallisto.temp33_1/abundance.h5

This just re-runs kallisto_quant - the final step.

BASH

$ rm -v trimmed/temp33_*.fq
$ snakemake -j1 -p kallisto.temp33_1/abundance.h5

“Nothing to be done” - some intermediate output is missing but Snakemake already has the file you are telling it to make, so it doesn’t worry.

BASH

$ touch transcriptome/*.fa.gz
$ snakemake -j1 -p kallisto.temp33_1/abundance.h5

The touch command is a standard Linux command which sets the timestamp of the file, so now the transcriptome looks to Snakemake as if it was just modified.

Snakemake sees that one of the input files used in the process of producing kallisto.temp33_1/abundance.h5 is newer than the existing output file, so it needs to run the kallisto index and kallisto quant steps again. Of course, the kallisto quant step needs the trimmed reads which we deleted earlier, so now the trimming step is re-run also.

Explicitly telling Snakemake what to re-run

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The default timestamp-based logic is really useful when you want to:

1. Change or add some inputs to an existing analysis without re-processing everything
2. Continue running a workflow that failed part-way

In most cases you can also rely on Snakemake to detect when you have edited a rule, but sometimes you need to be explicit, for example if you have updated an external script or changed a setting which Snakemake doesn’t see.

The -R flag allows you to explicitly tell Snakemake that a rule has changed and that all outputs from that rule need to be re-evaluated.

BASH

$ snakemake -j1 -R trimreads -p kallisto.temp33_1/abundance.h5

Note on -R

Due to a quirk of the way Snakemake parses command-line options, you need to make sure there are options after the -R ..., before the list of target outputs. If you don’t do this, Snakemake will think that the target files are instead items to add to the -R list, and then when building the DAG it will just try to run the default rule.

The easiest way is to put the -p flag before the target outputs. Then you can list multiple rules to re-run, and also multiple targets, and Snakemake can tell which is which.

BASH

$ snakemake -j1 -R trimreads kallisto_index -p kallisto.temp33_1/abundance.h5 kallisto.temp33_2/abundance.h5

The reason for using the -p flag specifically is that you pretty much always want this option.

The -f flag specifies that the target outputs named on the command line should always be regenerated, so you can use this to explicitly re-make specific files.

BASH

$ snakemake -j1 -f -p kallisto.temp33_1/abundance.h5

This always re-runs kallisto_quant, regardless of whether the output file is there already. For all intermediate outputs, Snakemake applies the default timestamp-based logic. Contrast with -F which runs the entire DAG every time.

Visualising the DAG

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Snakemake can draw a picture of the DAG for you, if you run it like this:

BASH

$ snakemake -f --dag kallisto.etoh60_1/abundance.h5 | gm display -

Using the --dag option implicitly activates the -n (dry-run) option so that Snakemake will not actually run any jobs, it will just print the DAG and stop. Snakemake prints the DAG in a text format so we use the gm command to make this into a picture and show it on the screen.

Note on gm display

The gm command is provided by the GraphicsMagick toolkit. On systems where gm will not display an image directly, you may instead save it to a PNG file. You will need the dot program from the GraphViz package installed.

BASH

$ snakemake -f --dag kallisto.etoh60_1/abundance.h5 | dot -Tpng > dag.png

The boxes drawn with dotted lines indicate steps that are not to be run, as the output files are already present and newer than the input files.

Visualising the effect of the -R and -f flags

Run kallisto_quant on the first of the etoh60 samples, then use the --dag option as shown above to check:

1. How many jobs will run if you ask again to create this output with no -f, -F or -R options?

2. How many if you use the -f option?

3. How many if you use the -R trimreads option?

4. How many if you edit the Snakefile so that the qual_threshold for trimreads is “22”, rather than “20”?

Show me the solution

This is a way to make the Kallisto result in the first place:

BASH

$ snakemake -j1 -p kallisto.etoh60_1/abundance.h5

1. This command should show four boxes, but all are dotted so no jobs are actually to be run.

BASH

$ snakemake --dag kallisto.etoh60_1/abundance.h5 | gm display -

2. The -f flag re-runs only the job to create the output file, so in this case one box is solid, and only that job will run.

3. With -R trimreads, the two trimreads jobs will re-run, and Snakemake sees that this also requires re-running kallisto_quant, so the answer is 3.

If you see a message like the one below, it’s because you need to put an option after trimreads or else Snakemake gets confused about what are parameters of -R, and what things are targets.

ERROR

WorkflowError:
Target rules may not contain wildcards.

4. Editing the Snakefile has the same effect as forcing the trimreads rule to re-run, so again there will be three jobs to be run from the DAG.

With older versions of Snakemake this would not be auto-detected, and in fact you can see this behaviour if you remove the hidden .snakemake directory. Now Snakemake has no memory of the rule change so it will not re-run any jobs unless explicitly told to.

Removing files to trigger reprocessing

In general, getting Snakemake to re-run things by removing files is a bad idea, because it’s easy to forget about intermediate files that actually contain stale results and need to be updated. Using the -R flag is simpler and more reliable. If in doubt, and if it will not be too time consuming, keep it simple and just use -F to run the whole workflow from scratch.

For the opposite case where you want to avoid re-running particular steps, see the ‑‑touch option of Snakemake mentioned later in the course.

Key Points

• A job in Snakemake is a rule plus wildcard values (determined by working back from the requested output)
• Snakemake plans its work by arranging all the jobs into a DAG (directed acyclic graph)
• If output files already exist, Snakemake can skip parts of the DAG
• Snakemake compares file timestamps and a log of previous runs to determine what need regenerating