TransDecoder - C.bairdi Transcriptome v4.0 on Mox

2021
Tanner Crab RNAseq
Author

Sam White

Published

March 17, 2021

Began annotation of cbai_transcriptome_v4.0.fasta Trinity de novo assembly from 20210317] using TransDecoder on Mox. This will be used as a component of Trinotate annotation downstream.

SBATCH script (GitHub):

#!/bin/bash
## Job Name
#SBATCH --job-name=20210317_cbai_transdecoder_transcriptome_v4.0
## Allocation Definition
#SBATCH --account=coenv
#SBATCH --partition=coenv
## Resources
## Nodes
#SBATCH --nodes=1
## Walltime (days-hours:minutes:seconds format)
#SBATCH --time=8-00:00:00
## Memory per node
#SBATCH --mem=120G
##turn on e-mail notification
#SBATCH --mail-type=ALL
#SBATCH --mail-user=samwhite@uw.edu
## Specify the working directory for this job
#SBATCH --chdir=/gscratch/scrubbed/samwhite/outputs/20210317_cbai_transdecoder_transcriptome_v4.0


# Exit script if a command fails
set -e

# Load Python Mox module for Python module availability

module load intel-python3_2017

# Set workind directory as current directory
wd="$(pwd)"


# Set input file locations
trinity_fasta="/gscratch/srlab/sam/data/C_bairdi/transcriptomes/cbai_transcriptome_v4.0.fasta"
trinity_gene_map="/gscratch/srlab/sam/data/C_bairdi/transcriptomes/cbai_transcriptome_v4.0.fasta.gene_trans_map"


# Capture trinity file name
trinity_fasta_name=${trinity_fasta##*/}



# Paths to input/output files
blastp_out_dir="${wd}/blastp_out"
transdecoder_out_dir="${wd}/${trinity_fasta_name}.transdecoder_dir"
pfam_out_dir="${wd}/pfam_out"
blastp_out="${blastp_out_dir}/${trinity_fasta_name}.blastp.outfmt6"
pfam_out="${pfam_out_dir}/${trinity_fasta_name}.pfam.domtblout"
lORFs_pep="${transdecoder_out_dir}/longest_orfs.pep"
pfam_db="/gscratch/srlab/programs/Trinotate-v3.1.1/admin/Pfam-A.hmm"
sp_db="/gscratch/srlab/programs/Trinotate-v3.1.1/admin/uniprot_sprot.pep"



# Paths to programs
blast_dir="/gscratch/srlab/programs/ncbi-blast-2.8.1+/bin"
blastp="${blast_dir}/blastp"
hmmer_dir="/gscratch/srlab/programs/hmmer-3.2.1/src"
hmmscan="${hmmer_dir}/hmmscan"
transdecoder_dir="/gscratch/srlab/programs/TransDecoder-v5.5.0"
transdecoder_lORFs="${transdecoder_dir}/TransDecoder.LongOrfs"
transdecoder_predict="${transdecoder_dir}/TransDecoder.Predict"

# Capture FastA MD5 checksum for future reference
md5sum "${trinity_fasta}" >> "${trinity_fasta_name}".checksum.md5

# Make output directories
mkdir "${blastp_out_dir}"
mkdir "${pfam_out_dir}"

# Extract long open reading frames
"${transdecoder_lORFs}" \
--gene_trans_map "${trinity_gene_map}" \
-t "${trinity_fasta}"

# Run blastp on long ORFs
"${blastp}" \
-query "${lORFs_pep}" \
-db "${sp_db}" \
-max_target_seqs 1 \
-outfmt 6 \
-evalue 1e-5 \
-num_threads 28 \
> "${blastp_out}"

# Run pfam search
"${hmmscan}" \
--cpu 28 \
--domtblout "${pfam_out}" \
"${pfam_db}" \
"${lORFs_pep}"

# Run Transdecoder with blastp and Pfam results
"${transdecoder_predict}" \
-t "${trinity_fasta}" \
--retain_pfam_hits "${pfam_out}" \
--retain_blastp_hits "${blastp_out}"

# Document programs in PATH (primarily for program version ID)
{
date
echo ""
echo "System PATH for $SLURM_JOB_ID"
echo ""
printf "%0.s-" {1..10}
echo "${PATH}" | tr : \\n
} >> system_path.log

RESULTS

Runtime was just under 8hrs:

TransDecoder runtime for cbai_transcriptome_v4.0 on Mox

Output folder: