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Troubleshooting

Symptom Cause Fix
igblastn: command not found scripts/install.sh not run bash scripts/install.sh
igblastn: Bad CPU type in executable x86-64 binary on Apple Silicon without Rosetta softwareupdate --install-rosetta, or build IgBLAST for arm64 (bioconda ships an osx-arm64 package)
Germline annotation database ... could not be found in [internal_data] IGDATA not set. IgBLAST resolves internal_data from it, falling back to the working directory Every call site sets IGDATA automatically; if running IgBLAST by hand, set IGDATA=geneGUI/data
DefineClones.py: command not found changeo/presto console scripts not on PATH Confirm python3 -m pip install --user -r backend/requirements.txt succeeded. The app asks Python where its scripts live; override a single tool with e.g. AUTOAB_DEFINECLONES, or prepend a directory with BCR_EXTRA_PATH.
Backend never starts, no output No Python on the machine has changeo/presto The app probes candidates and needs one that can import changeo, presto. Set AUTOAB_PYTHON=/path/to/python3.
R step fails with "there is no package called 'alakazam'" R dependencies missing install.packages(c("alakazam","shazam","ape"))
Rscript not found R not on PATH Set AUTOAB_RSCRIPT=/path/to/Rscript
Clone counts differ between runs on the same data Expected. The default GMM threshold estimate is not deterministic Pin the threshold by typing the same value when prompted, or set AUTOAB_THRESHOLD_METHOD=density. See VALIDATION.md.
Pipeline reports success but per-timepoint results are empty Timepoint mapping mismatch Fixed; see VALIDATION.md. If it reappears, check that timepoint_mapping.json keys match the FASTA basenames.
No CoV-AbDab matches Database CSV missing Confirm geneGUI/data/internal_data/CoV-AbDab_080224.csv exists
First analysis is very slow BLAST indexes being built from the reference FASTAs One-off; cached under geneGUI/data/Database-Files/ afterwards

Environment variables

Variable Effect
AUTOAB_PYTHON Python interpreter to run the pipeline with, skipping auto-detection
AUTOAB_RSCRIPT Path to Rscript
AUTOAB_MAKEDB, AUTOAB_DEFINECLONES, AUTOAB_CREATEGERMLINES, AUTOAB_BUILDTREES Explicit paths to individual Change-O console scripts
AUTOAB_IGBLASTN, AUTOAB_MAKEBLASTDB, AUTOAB_BLASTN Explicit paths to individual BLAST binaries
AUTOAB_THRESHOLD_METHOD density selects the deterministic kernel-density threshold instead of the default gamma-gamma GMM
BCR_EXTRA_PATH Prepended to PATH when the app spawns Python
BCR_IGHC_DB_PATH Path to a custom IGHC FASTA for the isotype step, instead of the bundled one

Platform notes

The IgBLAST binaries fetched by scripts/install.sh are x86-64 on both macOS and Linux; NCBI does not publish an arm64 macOS build for 1.22.0. On Apple Silicon they run under Rosetta 2, which is what the validation runs used. For a native arm64 build, install IgBLAST from bioconda (osx-arm64 is available) and copy igblastn, igblastp and makeblastdb into geneGUI/bin/.

Running the pipeline without the GUI

backend/pipeline_runner.py speaks newline-delimited JSON on stdin/stdout, so it can be driven directly. Send one config line:

{"action":"run","config":{"fasta_dir":"...","species":"mouse","database_type":"IMGT",
 "output_dir":"...","backend_dir":"...","clone_mode":"gene","linkage_method":"average"}}

then read messages of type progress, log, result and complete. When a threshold_request arrives, reply with {"type":"threshold_response","thresholds":{"<label>":<value>}} (or {"type":"threshold_response","value":<float>} in single-cohort mode).