LLMs-Universal-Life-Science-and-Clinical-Skills- mixcr-analysis

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name: bio-tcr-bcr-analysis-mixcr-analysis description: Perform V(D)J alignment and clonotype assembly from TCR-seq or BCR-seq data using MiXCR. Use when processing raw immune repertoire sequencing data to identify clonotypes and their frequencies. tool_type: cli primary_tool: MiXCR measurable_outcome: Execute skill workflow successfully with valid output within 15 minutes. allowed-tools:

  • read_file
  • run_shell_command

MiXCR Analysis

Complete Workflow (Recommended)

mixcr analyze generic-tcr-amplicon \
    --species human \
    --rna \
    --rigid-left-alignment-boundary \
    --floating-right-alignment-boundary C \
    input_R1.fastq.gz input_R2.fastq.gz \
    output_prefix

mixcr analyze 10x-vdj-tcr \
    input_R1.fastq.gz input_R2.fastq.gz \
    output_prefix

Step-by-Step Workflow

Step 1: Align Reads

mixcr align \
    --species human \
    --preset generic-tcr-amplicon-umi \
    input_R1.fastq.gz input_R2.fastq.gz \
    alignments.vdjca

mixcr align \
    --species human \
    --rna \
    -OallowPartialAlignments=true \
    input_R1.fastq.gz input_R2.fastq.gz \
    alignments.vdjca

Step 2: Refine and Assemble

mixcr refineTagsAndSort alignments.vdjca alignments_refined.vdjca

mixcr assemble alignments_refined.vdjca clones.clns

Step 3: Export Results

mixcr exportClones \
    --chains TRB \
    --preset full \
    clones.clns \
    clones.tsv

mixcr exportClones \
    --chains TRB \
    -cloneId -readCount -readFraction \
    -nFeature CDR3 -aaFeature CDR3 \
    -vGene -dGene -jGene \
    clones.clns \
    clones_custom.tsv

Preset Protocols

ProtocolUse Case
generic-tcr-amplicon
TCR amplicon sequencing
generic-bcr-amplicon
BCR amplicon sequencing
generic-tcr-amplicon-umi
TCR amplicon with UMIs
rnaseq-tcr
TCR extraction from bulk RNA-seq
rnaseq-bcr
BCR extraction from bulk RNA-seq
10x-vdj-tcr
10x Genomics TCR enrichment
10x-vdj-bcr
10x Genomics BCR enrichment
takara-human-tcr-v2
Takara SMARTer kit

Species Support

mixcr align --species human ...
mixcr align --species mmu ...

# Available: human, mmu, rat, rhesus, dog, pig, rabbit, chicken

Output Format

ColumnDescription
cloneIdUnique clone identifier
readCountNumber of reads
cloneFractionProportion of repertoire
nSeqCDR3Nucleotide CDR3 sequence
aaSeqCDR3Amino acid CDR3 sequence
allVHitsWithScoreV gene assignments
allDHitsWithScoreD gene assignments
allJHitsWithScoreJ gene assignments

Quality Metrics

mixcr exportReports alignments.vdjca

# Key metrics:
# - Successfully aligned reads (>80% is good)
# - CDR3 found (>70% of aligned)
# - Clonotype count (varies by sample type)

Parse MiXCR Output in Python

import pandas as pd

def load_mixcr_clones(filepath):
    df = pd.read_csv(filepath, sep='\t')
    df = df.rename(columns={
        'readCount': 'count',
        'cloneFraction': 'frequency',
        'aaSeqCDR3': 'cdr3_aa',
        'nSeqCDR3': 'cdr3_nt'
    })
    return df

Related Skills

  • vdjtools-analysis - Downstream diversity analysis
  • scirpy-analysis - Single-cell VDJ integration
  • repertoire-visualization - Visualize MiXCR output
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