A C implementation of the CaVEMan program. Uses an expectation maximisation approach to calling single base substitutions in paired data. Designed for use with a compute farm/cluster most steps in the program make use of an index parameter. The split step is designed to divide the genome into chunks of adjustable size to optimise for runtime/memory usage requirements.
- BWA Mapped, indexed, duplicate marked or removed bam/cram files, for both a normal and tumour sample
- Reference.fasta and index
- A one based bed style format file of regions to ignore during analysis (see specified format).
Optional inputs (will result in more accurate calls)
- Normal and tumour copy number files (see specified format).
- A normal contamination of tumour value
CaVEMan is executed in several distinct steps in the order listed below.
Generates a config file for use with the remaining CaVEMan steps (it'll save you a lot of time typing commandline args). Also generates a file named 'alg_bean' in the run directory.
bin/caveman setup || ./bin/setupCaveman Usage: caveman setup -t tum.bam -n norm.bam -r reference.fa.fai -g ignore_regions.tab -e tum_cn.bed -j norm_cn.bed [-f path] [-l path] [-a path] [-wzu] -t --tumour-bam [file] Location of tumour bam -n --normal-bam [file] Location of normal bam -r --reference-index [file] Location of reference fasta index -g --ignore-regions-file [file] Location of tsv ignore regions file Optional -c --config-file [file] File to write caveman run config file [default:'./caveman.cfg.ini'] -f --results-folder [file] Folder to write results [default:'./results'] -l --split-file [file] File to write list of split sections [default:'./splitList'] -a --alg-bean-file [file] Location to write alg-bean [default:'./alg_bean'] -w --include-smith-waterman Include SW mapped reads in the analysis -z --include-single-end Use single end reads for this analysis -u --include-duplicates Include reads marked as duplicates in the analysis -e --tumour-copy-no-file [file] Location of tumour copy number bed file (if the extension is not .bed the file will be treated as 1 based start). If no copy number file is supplied then the default cn of 2 will be used -j --normal-copy-no-file [file] Location of normal copy number bed file (if the extension is not .bed the file will be treated as 1 based start). If no copy number file is supplied then the default cn of 2 will be used -h --help Display this usage information.
Requires one job per entry in your reference.fasta.fai file. Each job creates a list of segments to be analysed, these are determined by total read count and do not include reads from ignored regions. The size of sections can be tuned using the -m, -c and -e parameters. Once all jobs complete successfully you will need to concatenate all split files into a single file with the name passed to the setup step in -f parameter (or splitList if you used the default).
bin/caveman split || ./bin/splitCaveman Usage: caveman split -i jobindex -f path [-c int] [-m int] [-e int] -f --config-file file Path to the config file produced by setup [default: 'caveman.cfg.ini']. -i --index int Job index (e.g. from $LSB_JOBINDEX) Optional -c --increment int Increment to use when deciding split sizes -m --max-read-count double Proportion of read-count to allow as a max in a split section -e --read-count int Guide for maximum read count in a section -h help Display this usage information.
Requires one job per entry in the merged split file. The parameter -i referes to a line in the merged split file. -a can be used to tune the size of section downloaded from bam at a time, this allows tuning of the memory footprint. The mstep will create a file for each job under the results folder (specified as a parameter in the setup step).
bin/caveman mstep || ./bin/mstepCaveman Usage: caveman mstep -f config.file -i jobindex [-m int] [-a int] -f --config-file file Path to the config file produced by setup [default: 'caveman.cfg.ini']. -i --index int Job index. Optional -a --split_size int Size of section to retrieve at a time from bam file. Allows memory footprint tuning [default:50000]. -m --min-base-qual int Minimum base quality to include in analysis [default:11] -h help Display this usage information.
Runs as a single job, merging all the cov_array files generated by the mstep into a file representing the profile of the whole genome. The resulting file is named cov_array and stored in the root run folder. Another file named prob_array is also created.
bin/caveman merge || ./bin/mergeCaveman Usage: caveman merge -f config_file [-c path] [-p path] -f --config-file file Path to the config file produced by setup. [default: 'caveman.cfg.ini'] Optional -c --covariate-file filename Location to write merged covariate array [default: covs_arr] -p --probabilities-file filename Location to write probability array [default: probs_arr] -h help Display this usage information.
The final step in calling variants using CaVEMan. As was the case with the mstep, a job per entry in the merged split list is required. Copy number (-e, -j), and normal contamination (-k) are required (See default settings for advice on obtaining results if you don't have these). Each job will create several files named according to the corresponding line in the merged split file. *_muts.vcf and *_snps.vcf are the calls above the given cutoffs for somatic and SNP probabilities alike. The *.no_analysis.bed file lists sections not analysed by caveman, due to being ignored or lacking coverage. Using the debugoption will output another file named *.dbg.vcf containing a line for every position in the genome that was analysed with read counts (as seen by CaVEMan) and top two probabilities calculated.
bin/caveman estep || ./bin/estepCaveman Usage: caveman estep -i jobindex [-f file] [-m int] [-k float] [-b float] [-p float] [-q float] [-x int] [-y int] [-c float] [-d float] [-a int] -i --index [int] Job index (e.g. from $LSB_JOBINDEX) Optional -f --config-file [file] Path to the config file produced by setup. [default:'caveman.cfg.ini'] -m --min-base-qual [int] Minimum base quality for inclusion of a read position [default:11] -c --prior-mut-probability [float] Prior somatic probability [default:0.000006] -d --prior-snp-probability [float] Prior germline mutant probability [default:0.000100] -k --normal-contamination [float] Normal contamination of tumour [default:0.100000] -b --reference-bias [float] Reference bias [default:0.950000] -p --mut-probability-cutoff [float] Minimum probability call for a somatic mutant position to be output [default:0.800000] -q --snp-probability-cutoff [float] Minimum probability call for a germline mutant position to be output [default:0.950000] -x --min-tum-coverage [int] Minimum tumour coverage for analysis of a position [default:1] -y --min-norm-coverage [int] Minimum normal coverage for analysis of a position [default:1] -a --split-size [int] Size of section to retrieve at a time from bam file. Allows memory footprint tuning [default:50000]. -s --debug Adds an extra output to a debug file. Every base analysed has an output -g --cov-file [file] File location of the covariate array. [default:'covs_arr'] -o --prob-file [file] File location of the prob array. [default:'probs_arr'] -v --species-assembly [string] Species assembly (eg 37/GRCh37), required if bam header SQ lines do not contain AS and SP information. -w --species [string] Species name (eg Human), required if bam header SQ lines do not contain AS and SP information. -n --normal-copy-number [int] Copy number to use when filling gaps in the normal copy number file [default:2]. -t --tumour-copy-number [int] Copy number to use when filling gaps in the tumour copy number file [default:2]. -l --normal-protocol [string] Normal protocol. Ideally this should match -r but not checked (WGS|WGX|RNA) [default:WGS]. -r --tumour-protocol [string] Tumour protocol. Ideally this should match -l but not checked (WGS|WGX|RNA) [default:WGS]. -P --normal-platform [string] Normal platform. Overrides the values retrieved from bam header. -T --tumour-platform [string] Tumour platform. Overrides the values retrieved from bam header. -M --max-copy-number [int] Maximum copy number permitted. If exceeded the copy number for the offending region will be set to this value. [default:10]. -h --help Display this usage information.
Copy number files are taken in a 1-based bed style tab separated format, or BED format if the suffix is .bed. Where the columns are chromosome,start,stop,copynumber(integer). A separate file is required for normal and tumour. Each file should have a copy number assigned for every region requested to be analysed (NB, CaVEMan set CN to 2 in regions where copy number is 0).
Example: 1 0 20000 2 2 0 2500 4 2 2501 500000 6
Ignored regions file
A 1-based bed style tab separated format file of regions to be ignored during analysis. An example might be regions known to have extreme depth of mapped reads through mismapping.
There may be cases where copy number is not available. In order to extract the best results it is advised to use copy number 2 in the normal and 5 in the tumour in combination with a normal contamination of 0.1 . This gives CaVEMan a broad range over which variants will be called compared to a copy number of 2 in the tumour.
Copyright (c) 2014-2015 Genome Research Ltd.
Author: Cancer Genome Project firstname.lastname@example.org
This file is part of CaVEMan.
CaVEMan is free software: you can redistribute it and/or modify it under the terms of the GNU Affero General Public License as published by the Free Software Foundation; either version 3 of the License, or (at your option) any later version.
This program is distributed in the hope that it will be useful, but WITHOUT ANY WARRANTY; without even the implied warranty of MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU Affero General Public License for more details.
You should have received a copy of the GNU Affero General Public License along with this program. If not, see http://www.gnu.org/licenses/.
- The usage of a range of years within a copyright statement contained within this distribution should be interpreted as being equivalent to a list of years including the first and last year specified and all consecutive years between them. For example, a copyright statement that reads ‘Copyright (c) 2005, 2007- 2009, 2011-2012’ should be interpreted as being identical to a statement that reads ‘Copyright (c) 2005, 2007, 2008, 2009, 2011, 2012’ and a copyright statement that reads ‘Copyright (c) 2005-2012’ should be interpreted as being identical to a statement that reads ‘Copyright (c) 2005, 2006, 2007, 2008, 2009, 2010, 2011, 2012’."