Bactopia Tool - ismapper

The ismapper module uses ISMapper to search for insertion sites in your samples.

Example Usage

bactopia --wf ismapper \
  --bactopia /path/to/your/bactopia/results \ 
  --include includes.txt  

Output Overview

Below is the default output structure for the ismapper tool. Where possible the file descriptions below were modified from a tools description.

ismapper/
├── <SAMPLE_NAME>
│   ├── <INSERTION_NAME>
│   │   └── <FASTA_ENTRY>
│   │       ├── <SAMPLE_NAME>_<FASTA_ENTRY>_{left_final,right_final}.fastq
│   │       ├── <SAMPLE_NAME>__<REFERENCE>_closest.bed
│   │       ├── <SAMPLE_NAME>__<REFERENCE>_intersect.bed
│   │       ├── <SAMPLE_NAME>__<REFERENCE>_table.txt
│   │       ├── <SAMPLE_NAME>_{left,right}_<REFERENCE>_finalcov.bed
│   │       ├── <SAMPLE_NAME>_{left,right}_<REFERENCE>_merged.sorted.bed
│   │       └── <SAMPLE_NAME>_{left,right}_<REFERENCE>_unpaired.bed
│   └── logs
│       └── ismapper
│           └── <INSERTION_NAME>
│               ├── <SAMPLE_NAME>.log
│               ├── nf-ismapper.{begin,err,log,out,run,sh,trace}
│               └── versions.yml
├── logs
│   └── custom_dumpsoftwareversions
│       ├── nf-custom_dumpsoftwareversions.{begin,err,log,out,run,sh,trace}
│       └── versions.yml
├── nf-reports
│   ├── ismapper-dag.dot
│   ├── ismapper-report.html
│   ├── ismapper-timeline.html
│   └── ismapper-trace.txt
├── software_versions.yml
└── software_versions_mqc.yml

Results

ISMapper

Below is a description of the per-sample results from ISMapper.

Extension	Description
_final.fastq	Sequences (FASTQ format) that mapped to the flanking regions of the IS query
_closest.bed	Merged regions that are close but do not overlap
_intersect.bed	An intersection of merged regions from the left and right flanks.
_table.txt	A detailed description of the IS query results.
_finalcov.bed	Contains information about the coverage of the IS query
_merged.sorted.bed	Merged overlapping regions that passed coverage cutoffs
_unpaired.bed	All unpaired mappings to the IS query

Audit Trail

Below are files that can assist you in understanding which parameters and program versions were used.

Logs

Each process that is executed will have a logs folder containing helpful files for you to review if the need ever arises.

Filename	Description
nf-<PROCESS_NAME>.begin	An empty file used to designate the process started
nf-<PROCESS_NAME>.err	Contains STDERR outputs from the process
nf-<PROCESS_NAME>.log	Contains both STDERR and STDOUT outputs from the process
nf-<PROCESS_NAME>.out	Contains STDOUT outputs from the process
nf-<PROCESS_NAME>.run	The script Nextflow uses to stage/unstage files and queue processes based on given profile
nf-<PROCESS_NAME>.sh	The script executed by bash for the process
nf-<PROCESS_NAME>.trace	The Nextflow Trace report for the process
versions.yml	A YAML formatted file with program versions

Nextflow Reports

These Nextflow reports provide great a great summary of your run. These can be used to optimize resource usage and estimate expected costs if using cloud platforms.

Filename	Description
ismapper-dag.dot	The Nextflow DAG visualisation
ismapper-report.html	The Nextflow Execution Report
ismapper-timeline.html	The Nextflow Timeline Report
ismapper-trace.txt	The Nextflow Trace report

Program Versions

At the end of each run, each of the versions.yml files are merged into the files below.

Filename	Description
software_versions.yml	A complete list of programs and versions used by each process
software_versions_mqc.yml	A complete list of programs and versions formatted for MultiQC

Parameters

Required Parameters

Define where the pipeline should find input data and save output data.

Parameter	Description	Default
--bactopia	The path to bactopia results to use as inputs

Filtering Parameters

Use these parameters to specify which samples to include or exclude.

Parameter	Description	Default
--include	A text file containing sample names (one per line) to include from the analysis
--exclude	A text file containing sample names (one per line) to exclude from the analysis

ISMapper Parameters

Parameter	Description	Default
--reference	Reference genome for typing against in GenBank format
--insertions	Multifasta file with insertion sequence(s) to be mapped to
--min_clip	Minimum size for softclipped region to be extracted from initial mapping	10
--max_clip	Maximum size for softclipped regions to be included	30
--cutoff	Minimum depth for mapped region to be kept in bed file	6
--novel_gap_size	Distance in base pairs between left and right flanks to be called a novel hit	15
--min_range	Minimum percent size of the gap to be called a known hit	0.9
--max_range	Maximum percent size of the gap to be called a known hit	1.1
--merging	Value for merging left and right hits in bed files together to simply calculation of closest and intersecting regions	100
--ismap_all	Switch on all alignment reporting for bwa	False
--ismap_minqual	Mapping quality score for bwa	30

Optional Parameters

These optional parameters can be useful in certain settings.

Parameter	Description	Default
--outdir	Base directory to write results to	./
--run_name	Name of the directory to hold results	bactopia
--skip_compression	Ouput files will not be compressed	False
--keep_all_files	Keeps all analysis files created	False

Max Job Request Parameters

Set the top limit for requested resources for any single job.

Parameter	Description	Default
--max_retry	Maximum times to retry a process before allowing it to fail.	3
--max_cpus	Maximum number of CPUs that can be requested for any single job.	4
--max_memory	Maximum amount of memory (in GB) that can be requested for any single job.	32
--max_time	Maximum amount of time (in minutes) that can be requested for any single job.	120
--max_downloads	Maximum number of samples to download at a time	3

Nextflow Configuration Parameters

Parameters to fine-tune your Nextflow setup.

Parameter	Description	Default
--nfconfig	A Nextflow compatible config file for custom profiles, loaded last and will overwrite existing variables if set.
--publish_dir_mode	Method used to save pipeline results to output directory.	copy
--infodir	Directory to keep pipeline Nextflow logs and reports.	${params.outdir}/pipeline_info
--force	Nextflow will overwrite existing output files.	False
--cleanup_workdir	After Bactopia is successfully executed, the work directory will be deleted.	False

Nextflow Profile Parameters

Parameters to fine-tune your Nextflow setup.

Parameter	Description	Default
--condadir	Directory to Nextflow should use for Conda environments
--registry	Docker registry to pull containers from.	dockerhub
--singularity_cache	Directory where remote Singularity images are stored.
--singularity_pull_docker_container	Instead of directly downloading Singularity images for use with Singularity, force the workflow to pull and convert Docker containers instead.
--force_rebuild	Force overwrite of existing pre-built environments.	False
--queue	Comma-separated name of the queue(s) to be used by a job scheduler (e.g. AWS Batch or SLURM)	general,high-memory
--cluster_opts	Additional options to pass to the executor. (e.g. SLURM: '--account=my_acct_name'
--disable_scratch	All intermediate files created on worker nodes of will be transferred to the head node.	False

Helpful Parameters

Uncommonly used parameters that might be useful.

Parameter	Description	Default
--monochrome_logs	Do not use coloured log outputs.
--nfdir	Print directory Nextflow has pulled Bactopia to
--sleep_time	The amount of time (seconds) Nextflow will wait after setting up datasets before execution.	5
--validate_params	Boolean whether to validate parameters against the schema at runtime	True
--help	Display help text.
--wf	Specify which workflow or Bactopia Tool to execute	bactopia
--list_wfs	List the available workflows and Bactopia Tools to use with '--wf'
--show_hidden_params	Show all params when using --help
--help_all	An alias for --help --show_hidden_params
--version	Display version text.

Citations

If you use Bactopia and ismapper in your analysis, please cite the following.

• Bactopia
  Petit III RA, Read TD Bactopia - a flexible pipeline for complete analysis of bacterial genomes. mSystems 5 (2020)

• ISMapper
  Hawkey J, Hamidian M, Wick RR, Edwards DJ, Billman-Jacobe H, Hall RM, Holt KE ISMapper: identifying transposase insertion sites in bacterial genomes from short read sequence data. BMC Genomics 16, 667 (2015)