First Steps: Episode 4¶

Episode	Topic
0	How can I install the tools?
1	How can I use the static data?
2	How can I distribute my jobs on the cluster (Slurm)?
3	How can I organize my jobs with Snakemake?
4	How can I combine Snakemake and Slurm?

In the last episodes we learned about distributing a job among the cluster nodes using sbatch and how to automate and parallelize our pipeline with Snakemake. We are lucky that those two powerful commands can be combined. What is the result? You will have an automated pipeline with Snakemake that uses sbatch to distribute jobs among the cluster nodes instead of running only the same node.

The best thing is that we can reuse our Snakefile as it is and just write a wrapper script to call Snakemake. We run the script and the magic will start.

First, create a new folder for this episode:

(first-steps) $ mkdir -p /fast/users/${USER}/work/tutorial/episode4/logs
(first-steps) $ pushd /fast/users/${USER}/work/tutorial/episode4

And copy the wrapper script to this folder as well as the Snakefile (you can also reuse the one with the adjustments from the previous episode):

(first-steps) $ cp /data/cephfs-1/work/projects/cubit/tutorial/skeletons/submit_snakejob.sh .
(first-steps) $ cp /data/cephfs-1/work/projects/cubit/tutorial/skeletons/Snakefile .
(first-steps) $ chmod u+w submit_snakejob.sh Snakefile

The Snakefile is already known to you but let me explain the wrapper script submit_snakejob.sh:

#!/bin/bash

# Set a name for the job (-J or --job-name).
#SBATCH --job-name=tutorial

# Set the file to write the stdout and stderr to (if -e is not set; -o or --output).
#SBATCH --output=logs/%x-%j.log

# Set the number of cores (-n or --ntasks).
#SBATCH --ntasks=2

# Force allocation of the two cores on ONE node.
#SBATCH --nodes=1

# Set the total memory. Units can be given in T|G|M|K.
#SBATCH --mem=1G

# Optionally, set the partition to be used (-p or --partition).
#SBATCH --partition=medium

# Set the expected running time of your job (-t or --time).
# Formats are MM:SS, HH:MM:SS, Days-HH, Days-HH:MM, Days-HH:MM:SS
#SBATCH --time=30:00


export TMPDIR=/fast/users/${USER}/scratch/tmp
export LOGDIR=logs/${SLURM_JOB_NAME}-${SLURM_JOB_ID}
mkdir -p $LOGDIR

eval "$($(which conda) shell.bash hook)"
conda activate first-steps

set -x

snakemake --profile=cubi-v1 -j 2 -k -p --restart-times=2

In the beginning you see the #SBATCH that introduces the parameters when you provide this script to sbatch as described in the second episode. Please make sure that the logs folder exists before starting the run! We then set and export the TMPDIR and LOGDIR variables. Note that LOGDIR has a subfolder named $SLURM_JOB_NAME-$SLURM_JOB_ID that will be created for you. Snakemake will store its logfiles for this very Snakemake run in this folder. The next new thing is set -x. This simply prints to the terminal every command that is executed within the script. This is useful for debugging.

Finally, the Snakemake call takes place. With the --profile option we define that Snakemake uses the Snakemake profile at /etc/xdg/snakemake/cubi-v1. The profile will take create appropriate calls to sbatch and interpret the following settings from your Snakemake rule:

threads: the number of threads to execute the job on
memory in megabytes or with a suffix of k, M, G, or T. You can specify EITHER
- resources.mem/resources.mem_mb: the memory to allocate for the whole job, OR
- resources.mem_per_thread: the memory to allocate for each thread.
resources.time: the running time of the rule, in a syntax supported by Slurm, e.g. HH:MM:SS or D-HH:MM:SS
resources.partition: the partition to submit your job into (Slurm will pick a fitting partition for you by default)
resources.nodes: the number of nodes to schedule your job on (defaults to 1 and you will want to keep that value unless you want to use MPI)

The other options to snakemake have the meaning:

-j 2: run at most two jobs at the same time
-k: keep going even if a rule execution fails
-p: print the executed shell commands
--restart-times=2: restart failing jobs up to two times

It is now time to update your Snakefile such that it actually specifies the resources mentioned above:

rule all:
    input:
        'snps/test.vcf',
        'structural_variants/test.vcf'

rule alignment:
    input:
        '/data/cephfs-1/work/projects/cubit/tutorial/input/{id}_R1.fq.gz',
        '/data/cephfs-1/work/projects/cubit/tutorial/input/{id}_R2.fq.gz',
    output:
        bam='alignment/{id}.bam',
        bai='alignment/{id}.bam.bai',
    threads: 8
    resources:
        mem='8G',
        time='12:00:00',
    shell:
        r"""
        export TMPDIR=/fast/users/${{USER}}/scratch/tmp
        mkdir -p ${{TMPDIR}}

        BWAREF=/data/cephfs-1/work/projects/cubit/current/static_data/precomputed/BWA/0.7.17/GRCh37/g1k_phase1/human_g1k_v37.fasta

        bwa mem -t 8 \
            -R "@RG\tID:FLOWCELL.LANE\tPL:ILLUMINA\tLB:test\tSM:PA01" \
            ${{BWAREF}} \
            {input} \
        | samtools view -b \
        | samtools sort -O BAM -T ${{TMPDIR}} -o {output.bam}

        samtools index {output.bam}
        """

rule structural_variants:
    input:
        'alignment/{id}.bam'
    output:
        'structural_variants/{id}.vcf'
    threads: 1
    resources:
        mem='4G',
        time='2-00:00:00',
    shell:
        r"""
        REF=/data/cephfs-1/work/projects/cubit/current/static_data/reference/GRCh37/g1k_phase1/human_g1k_v37.fasta

        delly call -o {output} -g ${{REF}} {input}
        """

def snps_mem(wildcards, attempt):
    mem = 2 * attempt
    return '%dG' % mem

rule snps:
    input:
        'alignment/{id}.bam'
    output:
        'snps/{id}.vcf'
    threads: 1
    resources:
        mem=snps_mem,
        time='04:00:00',
    shell:
        r"""
        REF=/data/cephfs-1/work/projects/cubit/current/static_data/reference/GRCh37/g1k_phase1/human_g1k_v37.fasta

        gatk HaplotypeCaller \
            -R ${{REF}} \
            -I {input} \
            -ploidy 2 \
            -O {output}
        """

We thus configure the resource consumption of the rules as follows:

alignment with 8 threads and up to 8GB of memory in total with a running time of up to 12 hours,
structural_variants with one thread and up to 4GB of memory in with a running time of up to 2 days,
snps with one thread and running up to four hours. Instead of passing a static amount of memory, we pass a resource callable. The attempt parameter will be passed a value of 1 on the initial invocation. If variant calling with the GATK HaplotypeCaller fails then it will retry and attempt will have an incremented value on each invocation (2 on the first retry and so on). Thus, we try to do small variant calling with 2, 4, 6, and 8 GB.

Finally, run the script:

(first-steps) $ sbatch submit_snakejob.sh

If you watch squeue --me now, you will see that the jobs are distributed to the system:

(first-steps) $ squeue --me

Please refer to the Snakemake documentation for more details on using Snakemake, in particular how to use the cluster configuration on how to specify the resource requirements on a per-rule base.