10x Genomics
Chromium Single Cell CNV

Cell Ranger DNA1.0, printed on 12/21/2024

Pipestance Structure

The pipeline output directory, described in Understanding Output, contains all of the data produced by one invocation of a pipeline (a pipestance) as well as rich metadata describing the characteristics of each stage. This directory contains a specific structure that is used by the Martian pipeline framework to track the state of the pipeline as execution proceeds.

Pipeline Structure

Cell Ranger DNA's notion of a pipeline is very flexible in that a pipeline can be composed of stages that run stage code or sub-pipelines that may themselves contain stages or sub-pipelines.

Cell Ranger DNA pipelines follow the convention that stages are named with verbs (e.g., TRIM_READS, MARK_DUPLICATES, DETECT_BREAKPOINTS) and sub-pipelines are named with nouns and prefixed with an underscore (e.g., _BCSORTER). Each stage runs in its own directory bearing its name, and each stage's directory is contained within its parent pipeline's directory.

For example, the cellranger-dna mkfastq pipeline has the following process graph:

where

• MAKE_FASTQS_CS is the top-level pipeline stage
• MAKE_FASTQS is a sub-pipeline contained in MAKE_FASTQS_CS
• PREPARE_SAMPLESHEET, BCL2FASTQ_WITH_SAMPLESHEET, MAKE_QC_SUMMARY, and MERGE_FASTQS_BY_LANE_SAMPLE are stages contained in the MAKE_FASTQS sub-pipeline.
• MAKE_FASTQS_PREFLIGHT and MAKE_FASTQS_PREFLIGHT_LOCAL are preflight stages, which validate inputs prior to running the other stages. These also belong to MAKE_FASTQS, but have no connections to other stages because they don't produce any outputs.

Directory Structure

Every pipestance operates wholly inside of its pipeline output directory. When the pipestance completes, this pipestance output directory contains three outputs: metadata files, the pipestance output file directory, and the top-level pipeline stage directory.

• Metadata files are files prefixed with an underscore (_) and usually contain unstructured text or JSON-encoded arrays and hashes.
• The pipestance output file directory is a directory called outs/ that contains the pipestance's output files.
• The top-level pipeline stage directory is a directory named according to the top-level pipeline stage that contains the child stage directories that compose this pipestance.

The top-level pipeline stage directory is a stage directory that contains any number of child stage directories as well as one stage output directory for each fork run by that stage. There are four possible top-level pipeline stages:

• MAKE_FASTQS_CS for cellranger-dna mkfastq
• CNV_CALLER_SINGLECELL_CS for cellranger-dna cnv

All of the Cell Ranger DNA pipelines only contain single-fork stages, so there will only ever be a fork0 stage output directory within each stage directory. Chunk output directories are a subset of stage output directories that additionally contain runtime information specific to the job or process being run by that chunk (e.g., a process ID or cluster job ID).

For example, the Cell Ranger DNA mkfastq pipeline's pipeline output directory contains the following directory structure:

Commonly Generated Metadata

The metadata contained in the pipeline output directory includes

Stage directories contain stage output directories, stage output files, and the stage directories of any child stages or pipelines.

Stage output directories typically contain:

Chunk output directories are a subset of stage output directories that, in addition to the aforementioned stage output, may contain:

These metadata files should be treated as read-only, and altering the contents of metadata files is not recommended.

Navigating Pipestances

Pipestance output directories can demonstrate very complicated structures, and re-attaching the Cell Ranger DNA UI is the easiest way to quickly navigate to a pipeline stage of interest and examine its metadata. In the absence of being able to access the UI, the standard find command can quickly return high-level information about a pipestance.

For example, to find the stages that resulted in the overall failure of a pipestance whose output directory is sample345/,

$ find sample345/ -name _errors
sample345/CNV_CALLER_SINGLECELL_CS/CNV_CALLER_SINGLECELL/_BREAKPOINT_PIPELINE/AGGREGATE_NODES/fork0/join-u82692d3c96/_errors

This tells us that the failed stage was AGGREGATE_NODES.

It can be helpful to view all _errors files' contents at once by piping to xargs cat:

$ find sample345/ -name _errors | xargs cat
 
Traceback (most recent call last):
  File "/home/jdoe/cellranger-dna-1.0.0/martian-cs/v3.0.0/adapters/python/martian_shell.py", line 541, in _main
    stage.main()
  File "/home/jdoe/cellranger-dna-1.0.0/martian-cs/v3.0.0/adapters/python/martian_shell.py", line 513, in main
    self._run(lambda: self._module.join(
  File "/home/jdoe/cellranger-dna-1.0.0/martian-cs/v3.0.0/adapters/python/martian_shell.py", line 476, in _run
    cmd()
  File "/home/jdoe/cellranger-dna-1.0.0/martian-cs/v3.0.0/adapters/python/martian_shell.py", line 514, in 
    args, outs, chunk_defs, chunk_outs))
  File "/home/jdoe/cellranger-dna-1.0.0/cellranger-dna-cs/1.0.0/mro/stages/cluster_breakpoints/aggregate_nodes/__init__.py", line 225, in join
    assert X.shape[1] == Y.shape[1]
AssertionError

In the above case, the error is an unhandled exception whose cause is not obvious; these sorts of failures should be reported to the 10x software support team for assistance with diagnosis.

Stages whose stage code run external binaries (for example, the ALIGN stage which runs BWA) often generate output to their stdout and stderr streams. These messages are captured in the _stdout and _stderr metadata files within the chunk output directories, and combining find and xargs cat to examine their contents can also assist with troubleshooting.