taxopipe 0.1.0

Bioinformatics workflow management tool for taxonomic profiling on HPC systems

TaxoPipe

A Python package for running taxonomic profiling workflows on HPC systems with conda environment support.

Overview

TaxoPipe simplifies the process of running taxonomic profiling workflows on High-Performance Computing (HPC) systems. It automates the following tasks:

• Connecting to HPC systems using SSH
• Uploading raw sequencing data
• Downloading and setting up reference databases
• Configuring and submitting Snakemake workflows with conda environment support
• Monitoring job progress and checking job status
• Retrieving results and abundance matrices

Installation

pip install taxopipe

Prerequisites

• SSH access to an HPC system
• Python 3.9 or higher
• Conda or Mamba installed on the HPC system (for environment activation)

Conda Environment Setup

TaxoPipe requires a conda environment on the HPC system. While TaxoPipe can automatically generate the environment file, you may want to manually create it before running the workflow:

1. Create a file named environment.yaml with the following content:

name: taxopipe_env
channels:
  - bioconda
  - conda-forge
  - defaults
dependencies:
  - python=3.9
  - snakemake-minimal=7.32.4
  - kraken2=2.1.3
  - bracken=2.8
  - krona=2.7.1
  - fastp=0.23.4
  - bowtie2=2.5.2
  - samtools=1.18
  - pandas=2.1.1
  - numpy=1.26.0
  - biopython=1.81
  - pip=23.3.1
  - pip:
    - scikit-bio==0.5.8

2. Create the conda environment on the HPC system:

# SSH into your HPC system
ssh [e-mail]

# Navigate to your working directory
cd /path/to/your/project

# Create the conda environment from the YAML file
conda env create -f environment.yaml

# Activate the environment
conda activate taxopipe_env

3. Once the environment is created, you can submit your TaxoPipe job using the --conda-env option to specify the environment name:

taxopipe hpc \
  -r /path/to/sample1_L001_R1.fastq.gz \
  [...other options...] \
  --conda-env taxopipe_env

Usage

Basic Commands

Submit a job

taxopipe hpc \
  -r /path/to/sample1_L001_R1.fastq.gz \
  -r /path/to/sample1_L001_R2.fastq.gz \
  -r /path/to/sample1_L002_R1.fastq.gz \
  -r /path/to/sample1_L002_R2.fastq.gz \
  -o /path/on/hpc/output \
  --kraken-db /path/on/hpc/kraken2_db \
  --corn-db /path/on/hpc/corn_db \
  --host hpc.example.edu \
  --username myuser \
  --password-auth

Example with real paths:

taxopipe hpc \
  -r /Users/username/Downloads/S1_L001_R1.fastq.gz \
  -r /Users/username/Downloads/S1_L001_R2.fastq.gz \
  -r /Users/username/Downloads/S1_L002_R1.fastq.gz \
  -r /Users/username/Downloads/S1_L002_R2.fastq.gz \
  -o /home/username/toxotest/output \
  --kraken-db /home/username/toxotest/Kraken2_DB \
  --corn-db /home/username/toxotest/corn_db \
  --host hpcr8o2rnp.uta.edu \
  --username username \
  --password-auth

Check job status

taxopipe status 12345 --host hpc.example.edu --username myuser --password-auth

Example with real job ID:

taxopipe status 23104 --host hpcr8o2rnp.uta.edu --username username --password-auth

Download results

taxopipe download \
  --output-dir /path/on/hpc/output \
  --local-dir ./results \
  --host hpc.example.edu \
  --username myuser

Download abundance matrix

To download just the abundance matrix file (the final result of the workflow):

taxopipe get-abundance \
  --output-dir /path/on/hpc/output \
  --local-dir ./results \
  --host hpc.example.edu \
  --username myuser \
  --password-auth

You can also wait for a specific job to complete before downloading:

taxopipe get-abundance \
  --output-dir /path/on/hpc/output \
  --local-dir ./results \
  --host hpc.example.edu \
  --username myuser \
  --password-auth \
  --job-id 23110

Command Options

HPC Command Options

• -r, --raw-files: Raw FASTQ files to process (use multiple -r flags for multiple files)
• -o, --output-dir: Output directory on the HPC system
• --kraken-db: Path to Kraken2 database on the HPC system
• --corn-db: Path to corn genome database for host removal
• --host: HPC hostname (default: uses SSH config)
• --username: HPC username (default: uses SSH config)
• --identity-file: Path to SSH identity file/private key (e.g., ~/.ssh/id_rsa_toxolib)
• --password-auth: Use password authentication instead of SSH keys
• --partition: HPC partition/queue to use (default: normal)
• --threads: Number of threads to request (default: 16)
• --memory: Memory to request (default: 200GB)
• --time: Time limit for the job (default: 48:00:00)
• --conda-env: Specify a custom conda environment name (default: taxopipe_env)
• --no-download-db: Skip downloading reference databases
• --no-upload-data: Skip uploading raw data files (use if files are already on HPC)
• --dry-run: Show what would be done without actually connecting to HPC
• --help: Show help message

Status Command Options

• job_id: The SLURM job ID to check (required)
• --host: HPC hostname
• --username: HPC username
• --identity-file: Path to SSH identity file (private key)
• --password-auth: Use password authentication instead of key-based

Download Command Options

• --output-dir: HPC output directory path (required)
• --local-dir: Local directory to save downloaded files (default: current directory)
• --host: HPC hostname
• --username: HPC username
• --identity-file: Path to SSH identity file (private key)
• --password-auth: Use password authentication instead of key-based
• --file-pattern: File pattern to download (default: *.csv)

Get-Abundance Command Options

• --output-dir: HPC output directory path (required)
• --local-dir: Local directory to save the abundance matrix file (default: current directory)
• --host: HPC hostname
• --username: HPC username
• --identity-file: Path to SSH identity file (private key)
• --password-auth: Use password authentication instead of key-based
• --job-id: Job ID to check before downloading (will wait for job completion)

Reference Databases

TaxoPipe can automatically download and set up the required reference databases:

• Kraken2 Database: Standard microbial database for taxonomic classification

  • Source: https://genome-idx.s3.amazonaws.com/kraken/k2_standard_20240112.tar.gz

• Corn Genome Database: Used for host removal

  • Source: https://glwasoilmetagenome.s3.us-east-1.amazonaws.com/corn_db.zip

Manual Database Setup

If you prefer to manually download and set up the reference databases:

1. Download and prepare the Kraken2 database:

# On your local machine
# Download the Kraken2 database
wget https://genome-idx.s3.amazonaws.com/kraken/k2_standard_20240112.tar.gz

# Extract the database
tar -xzf k2_standard_20240112.tar.gz -C k2_standard

# Transfer to HPC (replace with your HPC details)
scp -r k2_standard [e-mail]:/path/on/hpc/Kraken2_DB

2. Download and prepare the Corn database:

# On your local machine
# Download the Corn database
wget https://glwasoilmetagenome.s3.us-east-1.amazonaws.com/corn_db.zip

# Extract the database
unzip corn_db.zip -d corn_db

# Transfer to HPC (replace with your HPC details)
scp -r corn_db [e-mail]:/path/on/hpc/corn_db

3. Transfer your sequencing data:

# On your local machine
# Create a directory for your data on HPC
ssh [e-mail] "mkdir -p /path/on/hpc/output/Raw_Data"

# Transfer your FASTQ files
scp /path/to/local/sample_L001_R1.fastq.gz [e-mail]:/path/on/hpc/output/Raw_Data/
scp /path/to/local/sample_L001_R2.fastq.gz [e-mail]:/path/on/hpc/output/Raw_Data/
# Repeat for all your FASTQ files

4. Run TaxoPipe with the --no-upload-data flag:

taxopipe hpc \
  -r /path/to/sample1_L001_R1.fastq.gz \
  -r /path/to/sample1_L001_R2.fastq.gz \
  -o /path/on/hpc/output \
  --kraken-db /path/on/hpc/Kraken2_DB \
  --corn-db /path/on/hpc/corn_db \
  --host hpc.example.edu \
  --username myuser \
  --password-auth \
  --no-upload-data

Output Structure

output/
├── Raw_Data/                  # Raw input files
├── workflow/                  # Workflow files and scripts
│   ├── Snakefile              # Snakemake workflow definition
│   ├── config.yaml            # Workflow configuration
│   ├── environment.yaml       # Conda environment definition
│   ├── scripts/               # Helper scripts
│   │   └── create_abundance_matrix.py  # Script for abundance matrix creation
│   ├── submit_job.sh          # Job submission script
│   ├── taxopipe_[job_id].out  # SLURM output log file
│   └── taxopipe_[job_id].err  # SLURM error log file

## Job Log Files

TaxoPipe generates log files for each job submission. These log files contain important information about the job execution, including any errors or warnings.

### Log File Locations

The log files are created in the workflow directory with the following naming pattern:

- **Output log**: `[output_dir]/workflow/taxopipe_[job_id].out`
- **Error log**: `[output_dir]/workflow/taxopipe_[job_id].err`

For example, if your output directory is `/home/username/toxotest/output` and your job ID is `23109`, the log files would be located at:

/home/username/toxotest/output/workflow/taxopipe_23109.out /home/username/toxotest/output/workflow/taxopipe_23109.err

### Viewing Log Files

You can view the log files using the following commands:

```bash
# SSH into your HPC system
ssh [e-mail]

# View the output log
cat /path/to/output/workflow/taxopipe_[job_id].out

# View the error log
cat /path/to/output/workflow/taxopipe_[job_id].err

# Or use 'tail' to follow the logs in real-time
tail -f /path/to/output/workflow/taxopipe_[job_id].out

├── Results/ │ └── Taxonomic_Profiling/ │ ├── 1_DNA_Kraken2/ # Kraken2 classification results │ ├── 2_DNA_Bracken/ # Bracken abundance estimation │ ├── 3_DNA_Krona/ # Krona input files │ ├── 4_DNA_Krona_HTML/ # Krona visualizations │ └── 5_DNA_Relative_Abundance_Matrix_Python/ # Abundance matrices

## Workflow Features

- **Automatic Authentication**: Multiple authentication methods with fallback options
- **Network Resilience**: Handles network connectivity issues and provides offline mode support
- **Conda Environment Support**: Automatically activates conda environments on the HPC system
- **Flexible Environment Management**: Creates minimal conda environments with retry logic
- **Progress Tracking**: Detailed logging and progress bars for file transfers
- **Job Status Checking**: Monitors job status and provides detailed information
- **Result Retrieval**: Downloads abundance matrices and other result files
- **Raw Data Management**: Efficiently uploads raw data with duplicate detection

## License

MIT License