---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
layout: post
title: Metagenome Assemblies - P.generosa Water Samples Trimmed HiSeqX Data Using Megahit on Mox
date: '2019-03-27 20:30'
tags:
  - metagenomics
  - Panopea generosa
  - geoduck
  - mox
  - megahit
  - assembly
categories:
  - 2019
  - Miscellaneous
---

[I previously created a single metagenome assembly](https://robertslab.github.io/sams-notebook/posts/2019/2019-01-02-Metagenome-Assembly---P.generosa-Water-Sample-HiSeqX-Data-Using-Megahit/) using all of the sequencing data from this project.

Now, we want to compare the different water sample metagenomes to each other.

Here's how the sample names breakdown:

| Sample | Develomental Stage (days post-fertilization) | pH Treatment |
|--------|-------------------------|--------------|
| MG1    | 13                      | 8.2          |
| MG2    | 17                      | 8.2          |
| MG3    | 6                       | 7.1          |
| MG5    | 10                      | 8.2          |
| MG6    | 13                      | 7.1          |
| MG7    | 17                      | 7.1          |



SBATCH script (GitHub):

- [20190327_metagenomics_pgen_megahit.sh](https://github.com/RobertsLab/sams-notebook/blob/master/sbatch_scripts/20190327_metagenomics_pgen_megahit.sh)

```shell
#!/bin/bash
## Job Name
#SBATCH --job-name=megahit
## Allocation Definition
#SBATCH --account=coenv
#SBATCH --partition=coenv
## Resources
## Nodes
#SBATCH --nodes=1
## Walltime (days-hours:minutes:seconds format)
#SBATCH --time=25-00:00:00
## 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 --workdir=/gscratch/scrubbed/samwhite/outputs/20190327_metagenomics_pgen_megahit

# Load Python Mox module for Python module availability

module load intel-python3_2017

# Load Open MPI module for parallel, multi-node processing

module load icc_19-ompi_3.1.2

# SegFault fix?
export THREADS_DAEMON_MODEL=1

# Document programs in PATH (primarily for program version ID)

date >> system_path.log
echo "" >> system_path.log
echo "System PATH for $SLURM_JOB_ID" >> system_path.log
echo "" >> system_path.log
printf "%0.s-" {1..10} >> system_path.log
echo ${PATH} | tr : \\n >> system_path.log


# variables
wd=$(pwd)
fastq_dir=/gscratch/srlab/sam/data/metagenomics/P_generosa
megahit=/gscratch/srlab/programs/megahit_v1.1.4_LINUX_CPUONLY_x86_64-bin/megahit
bbmap_dir=/gscratch/srlab/programs/bbmap_38.34
samtools=/gscratch/srlab/programs/samtools-1.9/samtools
cpus=28

## Inititalize arrays
fastq_array_R1=()
fastq_array_R2=()
names_array=()

# Create array of fastq R1 files
for fastq in ${fastq_dir}/*R1*.gz
do
  fastq_array_R1+=(${fastq})
done

# Create array of fastq R2 files
for fastq in ${fastq_dir}/*R2*.gz
do
  fastq_array_R2+=(${fastq})
done

# Create array of sample names
## Uses parameter substitution to strip leading path from filename
## Uses awk to parse out sample name from filename
for R1_fastq in ${fastq_dir}/*R1*.gz
do
  names_array+=($(echo ${R1_fastq#${fastq_dir}} | awk -F"_" '{print $3 $4}'))
done

# Create list of fastq files used in analysis
## Uses parameter substitution to strip leading path from filename
for fastq in ${fastq_dir}*.gz
do
  echo ${fastq#${fastq_dir}} >> fastq.list.txt
done

# Loop through samples
for sample in ${!names_array[@]}
do
  sample_name=$(echo ${names_array[sample]})
  mkdir ${sample_name} && cd ${sample_name}
  # Run Megahit using paired-end reads
  ${megahit} \
  -1 ${fastq_array_R1[sample]} \
  -2 ${fastq_array_R2[sample]} \
  --num-cpu-threads ${cpus} \
  --out-prefix ${sample_name}

  # Create FastA index file
  ${samtools} faidx megahit_out/${sample_name}.contigs.fa

  # Determine coverage
  ## Align reads with BBmap BBwrap
  ${bbmap_dir}/bbwrap.sh \
  ref=megahit_out/${sample_name}.contigs.fa \
  in1=${fastq_array_R1[sample]} \
  in2=${fastq_array_R2[sample]} \
  out=${sample_name}.aln.sam.gz

  ## Output contig coverage
  ${bbmap_dir}/pileup.sh \
  in=${sample_name}.aln.sam.gz \
  out=${sample_name}.coverage.txt

  # Return to working directory
  cd ${wd}
done
```

---

# RESULTS

I'll assess these when I get Emma's individual assemblies and create a new post with that comparison.

Output folder:

- [20190327_metagenomics_pgen_megahit/](http://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit)

  - MG1 Output folder:

  - [20190327_metagenomics_pgen_megahit/MG1](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG1)

     - MG1 FastA:

       - [20190327_metagenomics_pgen_megahit/MG1/megahit_out/MG1.contigs.fa](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG1/megahit_out/MG1.contigs.fa)

   - MG2 Output folder:

     - [20190327_metagenomics_pgen_megahit/MG2](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG2)

       - MG2 FastA:

       - [20190327_metagenomics_pgen_megahit/MG1/megahit_out/MG2.contigs.fa](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG2/megahit_out/MG2.contigs.fa)

    - MG3 Output folder:

      - [20190327_metagenomics_pgen_megahit/MG3](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG3)

        - MG3 Fasta:

        - [20190327_metagenomics_pgen_megahit/MG3/megahit_out/MG3.contigs.fa](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG3/megahit_out/MG3.contigs.fa)

    - MG5 Output folder:

      - [20190327_metagenomics_pgen_megahit/MG5](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG5)

        - MG5 FastA:

          - [20190327_metagenomics_pgen_megahit/MG5/megahit_out/MG5.contigs.fa](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG5/megahit_out/MG5.contigs.fa)

    - MG6 Output folder:

      - [20190327_metagenomics_pgen_megahit/MG6](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG6)

        - MG6 FastA:

          - [20190327_metagenomics_pgen_megahit/MG6/megahit_out/MG6.contigs.fa](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG6/megahit_out/MG6.contigs.fa)

    - MG7 Output folder:

      - [20190327_metagenomics_pgen_megahit/MG7](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG7)

        - MG7 FastA:

          - [20190327_metagenomics_pgen_megahit/MG7/megahit_out/MG7.contigs.fa](https://gannet.fish.washington.edu/Atumefaciens/20190327_metagenomics_pgen_megahit/MG7/megahit_out/MG7.contigs.fa)