scallop on Biowulf

From the scallop repository:

Scallop is an accurate reference-based transcript assembler. Scallop features its high accuracy in assembling multi-exon transcripts as well as lowly expressed transcripts. Scallop achieves this improvement through a novel algorithm that can be proved preserving all phasing paths from paired-end reads, while also achieves both transcripts parsimony and coverage deviation minimization.

References:

Documentation
Important Notes

Interactive job
Interactive jobs should be used for debugging, graphics, or applications that cannot be run as batch jobs.

Allocate an interactive session and run the program. Sample session:

[user@biowulf]$ sinteractive --gres=lscratch:10 --mem=7g
salloc.exe: Pending job allocation 46116226
salloc.exe: job 46116226 queued and waiting for resources
salloc.exe: job 46116226 has been allocated resources
salloc.exe: Granted job allocation 46116226
salloc.exe: Waiting for resource configuration
salloc.exe: Nodes cn3144 are ready for job

[user@cn3144]$ module load scallop
[user@cn3144]$ cd /lscratch/$SLURM_JOB_ID
[user@cn3144]$ # copy some example data - paired end RNA-Seq (101nt) of human
      # skin aligned with STAR for about a quarter of chr8 including Myc.
      # this is stranded RNA-Seq data
[user@cn3144]$ cp $SCALLOP_TEST_DATA/ENCSR862RGX.bam .
[user@cn3144]$ ls -lh ENCSR862RGX.bam
-rw-r--r-- 1 user group 81M Apr 18 07:30 ENCSR862RGX.bam

[user@cn3144]$ # run scallop
[user@cn3144]$ scallop --library_type second --min_transcript_length 300 \
             -i ENCSR862RGX.bam -o ENCSR862RGX.gtf
command line: scallop --library_type second --min_transcript_length 300 -i ENCSR862RGX.bam -o ENCSR862RG X.gtf


Bundle 0: tid = 7, #hits = 93, #partial-exons = 22, range = chr8:101915822-102124299, orient = + (93, 0, 0)
process splice graph gene.0.0 type = 1, vertices = 3, edges = 0
process splice graph gene.0.1 type = 0, vertices = 5, edges = 4
process splice graph gene.0.2 type = 1, vertices = 3, edges = 0
process splice graph gene.0.3 type = 1, vertices = 3, edges = 0
[...snip...]
[user@cn3144]$ wc -l ENCSR862RGX.gtf
5057 ENCSR862RGX.gtf
[user@cn3144]$ egrep '"gene.1.5"' ENCSR862RGX.gtf | head -3
chr8    scallop transcript      102204502       102239040       1000    +       .       gene_id "gene.1.5"; transcript_id "gene.1.5.2"; RPKM "22.4072"; cov "1.9556";
chr8    scallop exon    102204502       102205959       1000    +       .       gene_id "gene.1.5"; transcript_id "gene.1.5.2"; exon "1";
chr8    scallop exon    102208095       102208285       1000    +       .       gene_id "gene.1.5"; transcript_id "gene.1.5.2"; exon "2";


[user@cn3144]$ exit
salloc.exe: Relinquishing job allocation 46116226
[user@biowulf]$

Example of scallop assembled transcripts for chr8:103,394,722-103,446,765. The scallop transcripts are shown in black. Gencode v24 annotation is shown in blue.

scallop example results

Batch job
Most jobs should be run as batch jobs.

Create a batch input file (e.g. scallop.sh), which uses the input file 'scallop.in'. For example:

#! /bin/bash

module load scallop/0.10.2 || exit 1
scallop --verbose 0 --library_type second \
    -i ENCSR862RGX.bam -o ENCSR862RGX.gtf

Submit this job using the Slurm sbatch command.

sbatch --cpus-per-task=2 --mem=7g scallop.sh
Swarm of Jobs
A swarm of jobs is an easy way to submit a set of independent commands requiring identical resources.

Create a swarmfile (e.g. scallop.swarm). For example:

scallop --verbose 0 --library_type second -i sample1.bam -o sample1.gtf
scallop --verbose 0 --library_type second -i sample2.bam -o sample2.gtf
scallop --verbose 0 --library_type second -i sample3.bam -o sample3.gtf

Submit this job using the swarm command.

swarm -f scallop.swarm -g 7 -t 2 --module scallop/0.10.2
where
-g # Number of Gigabytes of memory required for each process (1 line in the swarm command file)
-t # Number of threads/CPUs required for each process (1 line in the swarm command file).
--module scallop Loads the scallop module for each subjob in the swarm