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@sjhosui here are the requested Cell Ranger scripts we have used on O2. I've included the sbatch settings for slurm as well.
slurm config
#SBATCH --job-name=cellranger # Job name
#SBATCH --partition=medium # Partition name
#SBATCH --time=3-00:00 # Runtime in D-HH:MM format
#SBATCH --nodes=1 # Number of nodes (keep at 1)
#SBATCH --ntasks=1 # Number of tasks per node (keep at 1)
#SBATCH --cpus-per-task=16 # CPU cores requested per task (change for threaded jobs)
#SBATCH --mem=128G # Memory needed per node (total)
#SBATCH --error=jobid_%j.err # File to which STDERR will be written, including job ID
#SBATCH --output=jobid_%j.out # File to which STDOUT will be written, including job ID
#SBATCH --mail-type=ALL # Type of email notification (BEGIN, END, FAIL, ALL)
module load bcl2fastq/2.20.0.422
module load cellranger/2.1.1
localcores=$SLURM_CPUS_PER_TASK
localmem=128mkfastq
# Point to the Illumina BCL run directory
# Use a CSV file to specify the samples (see 10X website for examples)
# cellranger mkfastq --help
illumina_bcl_dir="AFCHK2VYCCXY"
csv="mkfastq.csv"
cellranger mkfastq \
--run="${illumina_bcl_dir}" \
--csv="${csv}" \
--localcores="${localcores}" \
--localmem="${localmem}"count
Note that --nosecondary here will skip the automatic downstream clustering.
fastqs="MKFASTQ_DIR"
transcriptome="/n/data1/cores/bcbio/10x_genomics/cellranger/refdata/XXX"
# Repeat this command per sample
cellranger count \
--id=WT-A-CML6022-p8 \
--sample=WT-A-CML6022-p8 \
--fastqs="${fastqs}" \
--transcriptome="${transcriptome}" \
--localcores="${localcores}" \
--localmem="${localmem}" \
--nosecondaryaggr
Aggregating Multiple Libraries
Run this after the per sample data processing has completed.
cellranger aggr \
--id="aggr" \
--csv="aggr.csv" \
--normalize="mapped" \
--localcores="${SLURM_CPUS_PER_TASK}" \
--localmem=128 \
--nosecondaryMetadata
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docsissues related to knowledgebase docsissues related to knowledgebase docssubcategory-scrnaseqsingle cell RNAseq pipelinesingle cell RNAseq pipelineurgentissues to solve as soon as possibleissues to solve as soon as possible