- What is nextflow?
- Installing nextflow
- Basic nextflow commands to run a workflow
Nextflow is a workflow framework that can be used by a bioinformatician to integrate all of her/his/their bash/python/perl/other scripts into a one cohesive pipeline that are portable, reproducible, scalable and checkpointed. Nextflow is its own DSL (Domain Specific Language) that extends a language called groovy which is an extension of Java that has language feature similarity to Python, Ruby and Smalltalk.
- Fast protyping – let’s you write a computational pipeline from smaller tasks
- Reproducibility – supports Docker and Singularity containers
- Portable – can run locally, Slurm, SGE, PBS, and cloud (Google, Kubernetes and AWS)
- Unified parallelism – can process chunks through the entire pipeline (QC -> align -> call snps)
- Continuous checkpoints – each chunk and process it goes through is checkpointed
- Stream oriented – promotes programming approach extending Unix pipes model.
1 2 3 4 5 6 7 8 9 10 11 12 13 Note: Oracle Java 8/9/10 is no longer available for public download (license change). First install and update brew from Terminal: /usr/bin/ruby -e "$(curl -fsSL https://raw.githubusercontent.com/Homebrew/install/master/install)" brew tap homebrew/cask-versions brew update NEW as of June 2019 To install the JDKs from AdoptOpenJDK: brew tap adoptopenjdk/openjdk ... then one of the following: brew cask install adoptopenjdk8 brew cask install adoptopenjdk9 brew cask install adoptopenjdk10 brew cask install adoptopenjdk11
Note: This will require that you have xcode installed (devtools) and the latest xcode may require that you have the latest operating system.
curl -s https://get.nextflow.io | bash
- Install Xcode from Mac App Store
brew install nextflow
- Install the Unix subsystem
- Install Java 8 or above
sudo apt get install openjdk-8-jdk
- Install Nextflow
curl -s https://get.nextflow.io | bash
Running a nextflow workflow
Nextflow has many commands but we are going to focus on the run, pull and drop commands
To execute any of these commands, we type
nextflow then the command
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Commands: clean Clean up project cache and work directories clone Clone a project into a folder config Print a project configuration console Launch Nextflow interactive console drop Delete the local copy of a project help Print the usage help for a command info Print project and system runtime information kuberun Execute a workflow in a Kubernetes cluster (experimental) list List all downloaded projects log Print executions log and runtime info pull Download or update a project run Execute a pipeline project self-update Update nextflow runtime to the latest available version view View project script file(s)
nextflow run can be used on a nextflow
main.nf script or can be run directly from github repository. Let’s try this using a nextflow blast script developed by Iowa State’s Genome Informatic’s Facility (ISUGIF) by calling it directly from the Github repo.
Command: Showing the usage statement
1 nextflow run isugifNF/blast --help
The output is a general usage statement on how to use this workflow to run blast.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 N E X T F L O W ~ version 20.07.1 Launching `isugifNF/blast` [backstabbing_franklin] - revision: 11f393fd09 [master] Usage: The typical command for running the pipeline is as follows: nextflow run main.nf --query QUERY.fasta --genome GENOME.fasta -profile local nextflow run main.nf --query QUERY.fasta --dbDir "blastDatabaseDirectory" --dbName "blastPrefixName" -profile local Mandatory arguments: --query Query fasta file of sequences you wish to BLAST --genome Genome from which BLAST databases will be generated or --query Query fasta file of sequences you wish to BLAST --dbDir BLAST database directory (full path required) --dbName Prefix name of the BLAST database -profile Configuration profile to use. Can use multiple (comma separated) Available: test, condo, ceres, local, nova Optional arguments: --outdir Output directory to place final BLAST output --outfmt Output format ['6 qseqid sseqid pident length mismatch gapopen qstart qend sstart send evalue bitscore qlen slen frames salltitles qcovs'] --options Additional options for BLAST command [-evalue 1e-3] --outfileName Prefix name for BLAST output [blastout] --threads Number of CPUs to use during blast job  --chunkSize Number of fasta records to use when splitting the query fasta file --app BLAST program to use [blastn;blastp,tblastn,blastx] --help This usage statement.
Command: running the test example
This nextflow script happens to have a test dataset to show it is functioning properly and is perfect to show how nextflow operates.
1 nextflow run isugifNF/blast -profile test,singularity
Side note, for users on Ceres HPC (SCINet) you can use the following command which will call
module load blast+ on Ceres.
1 nextflow run isugifNF/blast -profile test,ceres
The command will produce the following output which tells us that it was run locally and executed three processes
runBlast. The processes run were determined by the nextflow script from isugifNF/blast.
1 2 3 4 5 6 7 8 N E X T F L O W ~ version 20.07.1 Launching `isugifNF/blast` [fabulous_gautier] - revision: ef0477a498 [master] executor > local (12) executor > local (12) [62/1e6e19] process > software_check [100%] 1 of 1 ✔ [50/6038d9] process > runMakeBlastDB (1) [100%] 1 of 1 ✔ [32/cba660] process > runBlast (5) [100%] 10 of 10 ✔ WARN: Task runtime metrics are not reported when using macOS without a container engine
This script produces a work directory which is default for all nextflow scripts and an
out_dir directory which can be specified with
--outdir (see help above)
Here is a more comprehensive structure of the files generated. The key here is that all of the processes are run in the work directory and are given unique hash numbers. If a process fails, you can fix the error and rerun the previous command using
-resume and it will pick up where it failed.
-resume only has one
- not two as it is a nextflow command parameter and not a nextflow script parameter (as in –help above)
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 tree . |-- out_dir | |-- blast_output_combined.txt | |-- report.html | `-- timeline.html `-- work |-- 70 | `-- 8c3ea2bbba3d4f31647d6410a4eced | |-- blast.log | |-- blastout | `-- headtest.1.fasta -> /Users/severin/nextflow/nextflow-tutorial/blasttest/work/d3/4f9a6a126c4f0a3d6779f975ebcb7c/headtest.1.fasta |-- d3 | `-- 4f9a6a126c4f0a3d6779f975ebcb7c | `-- headtest.1.fasta `-- fe `-- b6bdf808a6ebfdd7ef94e702821ac0 `-- software_check.txt 8 directories, 8 files
Not all nextflow scripts will have a
--help function or a
-profile test. If they are written well with the intention of sharing they should.
Want to try it on your own dataset?
If you want to give it a go on your own dataset use the examples provided in the
--help usage statement
1. How to have the workflow create the blast database for me
Creates a blast database from the
--genome file and then performs blast using
1 nextflow run isugifNF/blast --query QUERY.fasta --genome GENOME.fasta -profile local
Note: you need to define QUERY.fasta and GENOME.fasta. Those files do not exist. Replace those files with files of your own.
2. How to have the workflow Use the blast database I provide
Uses the BLAST database specified by
--dbDir and then performs blast using
1 nextflow run isugifNF/blast --query QUERY.fasta --dbDir "blastDatabaseDirectory" --dbName "blastPrefixName" -profile local
Note: you need to define QUERY.fasta, blastDatabaseDirectory and GENOME.fasta. Those files do not exist. Replace those files with files of your own.
nextflow pull command
When you want the latest version of the nextflow script that you pulled from a github repository, you can execute the the
1 nextflow pull isugifNF/blast
How do you know you need to pull? You will get a message that looks like this.
1 NOTE: Your local project version looks outdated - a different revision is available in the remote repository [d1fcc0c445]
nextflow drop command
Sometimes the local copy of the nextflow github repo can’t
fast forward and it may be easier to remove the local copy and start fresh.
1 nextflow drop isugifNF/blast
Once it is dropped you can start again using
1 nextflow run isugifNF/blast
and it will repull the latest version.
Note: if the
nextflow drop command does not work you can manually remove this repository. It is located:
~/.nextflow/assets/isugifNF/blast or more generically
nextflow log command
Nextflow log will display all of the recent runs of nextflow in that directory.
1 nextflow log
A rather useful function of nextflow log is that you can use it to display all the actual commands that were run by a given workflow
1 nextflow log -f script lonely_minsky
How to resume a nextflow pipeline
If your pipeline doesn’t finish or errors at a specific process it can be restarted from where it left off using the
-resume nextflow parameter. Note: there is only one
- (dash) as it is a nextflow parameter and not a pipeline parameter that your nextflow script has defined.
Note: It is more reliable to use the unique name given for each run.
1 -resume lonely_minsky
Further Reading and Resources
- NextFlow curated list of tutorials
- NextFlow on YouTube
- NextFlow Patterns are examples of more complex functionality.
- Searching for a biocontainer
- nf-core github repo of nextflow workflows.
- nf-core modules