ngs-ai-agent 1.1.0

AI-powered automated NGS analysis pipeline

NGS AI Agent

AI-powered automated NGS analysis pipeline for Deep Mutational Scanning (DMS) experiments

NGS AI Agent is a comprehensive, intelligent pipeline that automates Next-Generation Sequencing (NGS) data analysis with AI-powered metadata interpretation and experimental design detection. It supports both amplicon-based and barcode-coupled DMS workflows with minimal user configuration.

🚀 Quick Start

# 1. Create conda environment from environment.yml
conda env create -f environment.yml
conda activate ai-ngs

# 2. Install the package
pip install ngs-ai-agent

# 3. Run analysis with your data
ngs-ai-agent run --input-dir /path/to/fastq/files --metadata experiment.csv --dry-run

✨ Features

🤖 AI-Powered Analysis

• Intelligent File Matching: Automatically matches FASTQ files to experimental metadata using AI
• Pipeline Type Detection: Automatically detects amplicon-based vs barcode-coupled DMS experiments
• Condition Classification: AI-powered classification of samples as input, output, mapping, or control
• Metadata Interpretation: Supports CSV, TSV, and Excel metadata files with flexible column naming

🧬 Comprehensive DMS Support

• Amplicon-Based DMS: Direct sequencing of amplicons for variant calling and fitness calculation
• Barcode-Coupled DMS: Long-read barcode-to-variant mapping with short-read barcode counting
• Paired-End Support: Full support for paired-end sequencing data
• Quality Control: Integrated QC metrics and visualization

🔧 Advanced Workflow Management

• Snakemake Integration: Robust workflow management with automatic dependency resolution
• Conda Environment: Automated environment setup and dependency management
• Multi-Core Processing: Efficient parallel processing with configurable core usage
• Dry Run Mode: Preview pipeline execution before running

📊 Rich Output and Visualization

• Interactive Reports: HTML reports with embedded visualizations
• Fitness Calculations: Comprehensive fitness score calculations and statistical analysis
• Variant Analysis: Detailed variant calling and annotation
• Publication-Ready Plots: High-quality figures for manuscripts

📋 Requirements

• Python: 3.8 or higher
• Conda: For environment management
• Ollama: For AI capabilities (optional but recommended)
• Memory: 8GB+ RAM recommended
• Storage: Varies by dataset size

🛠️ Installation

Prerequisites

1. Install Conda: Download and install Miniconda or Anaconda

2. Clone the Repository:

git clone https://github.com/your-org/ngs-ai-agent.git
cd ngs-ai-agent

Step 1: Create Conda Environment

# Create the conda environment with all bioinformatics dependencies
conda env create -f environment.yml

# Activate the environment
conda activate ai-ngs

Step 2: Install the Package

# Install from PyPI (recommended)
pip install ngs-ai-agent

# OR install from source for development
pip install -e .

🎯 Usage

Basic Usage

# Make sure you're in the activated conda environment
conda activate ai-ngs

# Run analysis with CSV metadata
ngs-ai-agent run \
  --input-dir /path/to/fastq/files \
  --reference /path/to/reference.fasta \
  --metadata experiment.csv \
  --outdir /path/to/results \
  --cores 8

# Dry run to preview pipeline
ngs-ai-agent run \
  --input-dir /path/to/fastq/files \
  --metadata experiment.csv \
  --dry-run

Advanced Usage

# High-performance run with many cores
ngs-ai-agent run \
  --input-dir /path/to/fastq/files \
  --reference /path/to/reference.fasta \
  --metadata experiment.xlsx \
  --outdir /path/to/results \
  --cores 32 

# Custom configuration
ngs-ai-agent run \
  --input-dir /path/to/fastq/files \
  --metadata experiment.csv \
  --config /path/to/custom/config.yaml

Command Options

Option	Short	Description	Required
--input-dir	-i	Directory containing FASTQ files	✅
--reference	-r	Reference genome FASTA file	❌
--metadata	-m	Experimental metadata file (CSV/TSV/Excel)	❌
--config	-c	Configuration file path	❌
--cores	-j	Number of cores to use	❌
--dry-run		Show what would be run without executing	❌
--outdir	-o	Override output/results directory	❌

📁 Input Data Format

FASTQ Files

• Supported formats: .fastq.gz, .fq.gz, .fastq, .fq
• Paired-end: Automatically detected and handled
• Naming: Flexible naming conventions supported

Metadata Files

The pipeline supports CSV, TSV, and Excel metadata files with the following columns:

Required Columns

• Sample identification: filename1, filename, sample_name, or similar
• Condition: condition, treatment, timepoint, or similar
• Replication: replication, rep, replicate, or similar

Optional Columns

• Paired-end: filename2, filename_r2, file_name_r2 for R2 files
• Library layout: library_layout (single/paired)
• Description: description, sample_description for AI analysis

Example Metadata (CSV)

filename1,filename2,condition,replication,description
Sample1_R1.fastq.gz,Sample1_R2.fastq.gz,input,1,pre-selection library
Sample2_R1.fastq.gz,Sample2_R2.fastq.gz,output,1,post-selection enriched
Sample3_R1.fastq.gz,Sample3_R2.fastq.gz,mapping,1,PacBio barcode mapping

🔬 Pipeline Types

Amplicon-Based DMS

• Use case: Direct sequencing of PCR amplicons
• Workflow: Quality control → Variant calling → Fitness calculation
• Output: Variant frequencies, fitness scores, statistical analysis

Barcode-Coupled DMS

• Use case: Barcode-based variant identification
• Workflow: Barcode mapping → Barcode counting → Fitness calculation
• Output: Barcode-to-variant maps, fitness scores, enrichment analysis

📊 Output Structure

results/
├── reports/
│   └── final_report.html          # Interactive HTML report
├── variants/
│   └── final_variants.csv         # Variant calling results
├── fitness/
│   └── fitness_scores.csv         # Fitness calculations
├── plots/
│   ├── summary_plots.html         # Interactive visualizations
│   └── publication_plots/         # High-resolution figures
└── logs/
    └── pipeline.log               # Detailed execution log

⚙️ Configuration

The pipeline uses YAML configuration files. Key settings include:

# AI Settings
ai:
  model: "qwen3-coder:latest"
  endpoint: "http://localhost:11434"

# Pipeline Settings
pipeline:
  threads: 8
  memory: "16G"

# Data Paths
data:
  raw: "data/raw"
  processed: "data/processed"
  results: "results"

🤖 AI Integration

NGS AI Agent integrates with Ollama for intelligent analysis:

Setup Ollama

# Install Ollama
curl -fsSL https://ollama.ai/install.sh | sh

# Pull the required model
ollama pull qwen3-coder:latest

# Start Ollama service
ollama serve

AI Capabilities

• File Matching: Intelligent matching of files to metadata
• Condition Detection: Automatic classification of experimental conditions
• Pipeline Selection: Smart detection of appropriate analysis workflow
• Quality Assessment: AI-powered quality control recommendations

🧪 Examples

Example 1: Amplicon-Based DMS

ngs-ai-agent run \
  --input-dir data/amplicon_experiment/ \
  --reference reference.fasta \
  --metadata amplicon_metadata.csv \
  --cores 16

Example 2: Barcode-Coupled DMS

ngs-ai-agent run \
  --input-dir data/barcode_experiment/ \
  --reference barcode_reference.gb \
  --metadata barcode_metadata.xlsx \
  --cores 32 \
  --outdir results/barcode_analysis

🐛 Troubleshooting

Common Issues

1. Ollama Service Not Running

# Start Ollama service
ollama serve

# Check if model is available
ollama list

2. Conda Environment Issues

# Recreate environment
conda env remove -n ai-ngs
conda env create -f environment.yml
conda activate ai-ngs

3. Permission Errors

# Install with user flag
pip install --user ngs-ai-agent

4. Memory Issues

# Reduce cores or increase available memory
ngs-ai-agent run --cores 4 --input-dir /path/to/data --metadata experiment.csv

Getting Help

# Show help
ngs-ai-agent --help
ngs-ai-agent run --help

# Check version
ngs-ai-agent --version

🤝 Contributing

We welcome contributions! Please see our Contributing Guidelines for details.

Development Setup

# Clone repository
git clone https://github.com/your-org/ngs-ai-agent.git
cd ngs-ai-agent

# Create conda environment
conda env create -f environment.yml
conda activate ai-ngs

# Install in development mode
pip install -e .

# Install development dependencies
pip install -e ".[dev]"

# Run tests
pytest

📄 License

This project is licensed under the MIT License - see the LICENSE file for details.

📚 Citation

If you use NGS AI Agent in your research, please cite:

@software{ngs_ai_agent,
  title={NGS AI Agent: AI-powered automated NGS analysis pipeline},
  author={NGS AI Agent Team},
  year={2024},
  url={https://github.com/your-org/ngs-ai-agent},
  license={MIT}
}

🔗 Links

• Documentation: https://ngs-ai-agent.readthedocs.io/
• PyPI Package: https://pypi.org/project/ngs-ai-agent/
• GitHub Repository: https://github.com/your-org/ngs-ai-agent
• Issue Tracker: https://github.com/your-org/ngs-ai-agent/issues

🙏 Acknowledgments

• Built with Snakemake for workflow management
• AI capabilities powered by Ollama
• Bioinformatics tools from Bioconda
• Visualization with Plotly and Matplotlib

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Made with ❤️ for the bioinformatics community