bio-workflows-tcr-pipeline

$npx mdskill add GPTomics/bioSkills/bio-workflows-tcr-pipeline

Analyzes TCR/BCR repertoire data from FASTQ to clonotype diversity metrics using MiXCR and VDJtools

  • Processes immune repertoire sequencing data from raw FASTQ files to diversity metrics
  • Relies on MiXCR for alignment and clonotype assembly, VDJtools for diversity analysis
  • Follows a predefined pipeline workflow with configurable parameters for bulk or single-cell data
  • Exports clonotype data and diversity statistics for downstream visualization and interpretation
SKILL.md
.github/skills/bio-workflows-tcr-pipelineView on GitHub ↗
---
name: bio-workflows-tcr-pipeline
description: End-to-end TCR/BCR repertoire analysis from FASTQ to clonotype diversity metrics. Use when analyzing immune repertoire sequencing data from bulk or single-cell experiments.
tool_type: cli
primary_tool: MiXCR
---

## Version Compatibility

Reference examples tested with: MiXCR 4.6+, VDJtools 1.2.1+

Before using code patterns, verify installed versions match. If versions differ:
- CLI: `<tool> --version` then `<tool> --help` to confirm flags

If code throws ImportError, AttributeError, or TypeError, introspect the installed
package and adapt the example to match the actual API rather than retrying.

# TCR/BCR Analysis Pipeline

**"Analyze my TCR/BCR repertoire sequencing data end-to-end"** → Orchestrate MiXCR clonotype extraction, VDJtools diversity/repertoire analysis, Immcantation SHM and lineage analysis, and visualization of V/J gene usage and clonal dynamics.

## Pipeline Overview

```
FASTQ → MiXCR align → Assemble → Export → VDJtools diversity → Visualization
```

## Step 1: MiXCR Processing

```bash
# Align reads to V(D)J segments
mixcr align -s hsa -p rna-seq \
    R1.fastq.gz R2.fastq.gz \
    aligned.vdjca

# Assemble clonotypes
mixcr assemble aligned.vdjca clones.clns

# Export
mixcr exportClones clones.clns clones.txt
```

## Step 2: VDJtools Analysis

```bash
# Convert to VDJtools format
vdjtools Convert -S mixcr clones.txt vdjtools/

# Diversity metrics
vdjtools CalcDiversityStats vdjtools/clones.txt diversity/

# Sample overlap
vdjtools CalcPairwiseDistances vdjtools/*.txt overlap/
```

## Step 3: Visualization

```bash
# Spectratype plot
vdjtools PlotFancySpectratype vdjtools/clones.txt spectra/

# V usage
vdjtools PlotFancyVJUsage vdjtools/clones.txt usage/
```

## QC Checkpoints

1. **After alignment**: Check V/J assignment rate (>70% typical)
2. **After assembly**: Verify clonotype count and coverage
3. **After diversity**: Compare metrics to expected range

## Related Skills

- tcr-bcr-analysis/mixcr-analysis - Detailed MiXCR usage
- tcr-bcr-analysis/vdjtools-analysis - Diversity metrics
- tcr-bcr-analysis/repertoire-visualization - Plots
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