STAR
STAR (Spliced Transcripts Alignment to a Reference) is a fast NGS read aligner for RNA-seq data.
License
Free to use and open source under GNU GPLv3.
Available
Puhti: 2.7.10a, 2.7.11a
Usage
The STAR commands listed below are activated by loading biokit module.
Before you can run the actual alignment job, you must index your fasta formatted reference genome. In Puhti the working copies of reference genome indexes, as well as any large files, should be stored to the /scatch directory.
For ease of use, set an environment variable to point to your /scratch directory. (Substitute correct path for the one used in example).
Create a directory for the reference genome index:
After that, the indexing can be done with command:
STAR --runMode genomeGenerate --genomeDir $SCRATCH/star-genome --genomeFastaFiles /path/to/genome/genome.fasta --runThreadN 2
Once the indexing is done, the actual mapping task can be launched. STAR will generate the mapping output using fixed file names. Because of that it is recommended that each STAR job is run in a new, empty directory. In Puhti you should create this new job directory to /scratch directory of your project. New directory called starjob1 can be created with command:
after that the actual mapping job can be launched with commands:
The default parameters STAR uses are typical for mapping 2x76 or 2x101 Illumina reads to the human genome.
In Puhti, all computing tasks should be executed as batch jobs. In batch jobs you can also utilize thread based parallelization. Below is a sample batch job file for STAR. The job uses six computing cores from a single computing node. The memory reservation is 24 GB. Note that you must change the --account setting to match you poject.
#!/bin/bash -l
#SBATCH --job-name=STAR
#SBATCH --output=STAR.stdout
#SBATCH --error=STAR.stderr
#SBATCH --partition=small
#SBATCH --ntasks=1
#SBATCH --nodes=1
#SBATCH --cpus-per-task=6
#SBATCH --account=project_1234567
#SBATCH --mem=24000
export SCRATCH=/scratch/project_12345/$USER
# calculate indexes. You don't need to recalculte the indexes if they already exist.
mkdir $SCRATCH/star-genome
STAR --runMode genomeGenerate --genomeDir $SCRATSCH/star-genome --genomeFastaFiles /path/to/genome/genome.fasta --runThreadN $SLURM_CPUS_PER_TASK
# Run the mapping task
STAR --genomeDir $SCRATCH/star-genome --readFilesIn my-reads.fastq --runThreadN $SLURM_CPUS_PER_TASK
The batch job script is launced with command sbatch. For example: