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All command line options

friedue edited this page Feb 14, 2014 · 28 revisions

deepTools version: 1.5.7 (February 2014)

WORK IN PROGRESS

Here, you will find all the options available for the command line (almost all of them are also available in Galaxy, perhaps named slightly different).

You can always see all available command-line options via --help: $ /deepTools/bin/bamCoverage --help

A typical deepTools command could look like this:

$ /deepTools/bin/bamCoverage --bam myAlignedReads.bam \
--outFileName myCoverageFile.bigWig \
--outFileFormat bigwig \
--fragmentLength 200 \
--ignoreDuplicates \
--scaleFactor 0.5

There are several types of options and parameters in the command. To tell a program to use a certain option (e.g. to ignore duplicate reads), you will have to give the option name preceded by two hyphens (e.g. --ignoreDuplicates). In the tables on this page, we try to list:

  • the option name as recognized by the program
  • the kind of value that is sometimes expected after the option name (see the annotated figure below)
  • a verbose explanation of what the option actually does

The texts here are adjusted for readability, they might not match the help text that you see in the command line word by word.

Table of Content

bamCorrelate

bamFingerprint

bamCoverage

bamCompare


bamCorrelate

bamCorrelate can be run in two modes: bins and BED-file.

A typical command would like this:

$ /deepTools/bin/bamCorrelate BED-file \
--BED myRegionsOfInterest.bed \
--bamfiles myAlignedReads_Sample1.bam myAlignedReads_Sample2.bam \
--plotFile correlation_plot.png \
--corMethod pearson

For details on bamCorrelate, see the tool details.

Mandatory arguments

Command Expected Input Explanation
--bamfiles FILENAMES List of indexed BAM files separated by space (default: None)
--plotFile FILENAME File name to save the file containing the heatmap of the correlation. The file ending will be used to determine the image format, for example, if correlation.pdf is provided, the heatmap will be saved in pdf format. Available file format options are: .png, .emf, .eps, .pdf and .svg. (default: None)
--corMethod {pearson, spearman} Choose the method for the correlation calculation (default: None)

bamCorrelate: optional arguments

only for BED-file mode:

| Command | Expected Input | Explanation | |:----:|:----:|:----| | --BED | FILENAME | If the comparison of read counts should be limited to certain regions, a BED file can be given. If this is the case, then the correlation is computed for the number of reads that overlap such regions. (default: None) |

only for bins mode:

| Command | Expected Input | Explanation | |:----:|:----:|:----| | --binSize | INT | Length in base pairs for a window used to sample the genome. (default: 10000) | | --numberOfSamples | INT | Number of random genome places to count reads for the correlation computation. (default: 1000000) |

Command Expected Input Explanation
--includeZeros If set, then zero counts that happen for all BAM files are included. The default behavior is to ignore those cases (default: False)
--region CHR:START:END Region of the genome to limit the operation to - this is useful when testing parameters to reduce the computing time. The format is chr:start:end, for example: --region chr10 or --region chr10:456700:891000. (default: None)
--numberOfProcessors INT Number of processors to use. Type "max/2" to use half the maximum number of processors or "max" to use all available processors. (default: max/2)
--verbose Set to see processing messages. (default: False)

bamCorrelate: optional arguments for processing of the reads

Command Expected Input Explanation
--fragmentLength INT Length of the average fragment size. Reads will be extended to match this length unless they are paired-end, in which case they will be extended to match the fragment length. If this value is set to the read length or smaller, the read will not be extended. Warning the fragment length affects the normalization to 1x (see --normalizeUsingSequencingDepth). The formula to normalize using the sequencing depth is genomeSize/(number of mapped reads * fragmentLength). NOTE: If the BAM files contain mated and unmated paired-end reads, unmated reads will be extended to match the --fragmentLength. (default: 200)
--doNotExtendPairedEnds If set, reads are not extended to match the fragment length reported in the BAM file, instead they will be extended to match the --fragmentLength. Default is to extend the reads if paired-end information is available. (default: False)
--ignoreDuplicates If set, reads that have the same orientation and start position will be considered only once. If reads are paired, the mate position also has to coincide to ignore a read. (default: False)
--minMappingQuality INT If set, only reads that have a mapping quality score higher than --minMappingQuality are considered. (default: None)

bamCorrelate: arguments for the heatmap display

Command Expected Input Explanation
--zMin NUMBER Minimum value for the heatmap intensities. If not specified the value is set automatically (default: None)
--zMax NUMBER Maximum value for the heatmap intensities. If not specified the value is set automatically (default: None)
--colorMap {Spectral, summer, coolwarm, Set1, Set2, Set3, Dark2, hot, RdPu, YlGnBu, RdYlBu, gist_stern, cool, gray, GnBu, gist_ncar, gist_rainbow, CMRmap, bone, RdYlGn, spring, terrain, PuBu, spectral, gist_yarg, BuGn, bwr, cubehelix, YlOrRd, Greens, PRGn, gist_heat, Paired, hsv, Pastel2, Pastel1, BuPu, copper, OrRd, brg, gnuplot2, jet, gist_earth, Oranges, PiYG, YlGn, Accent, gist_gray, flag, BrBG, Reds, RdGy, PuRd, Blues, Greys, autumn, pink, binary, winter, gnuplot, RdBu, prism, YlOrBr, rainbow, seismic, Purples, ocean, PuOr, PuBuGn, afmhot} Color map to use for the heatmap. For exemplary plots for all the color schemes, click here (default: Reds)

bamCorrelate: optional arguments concerning the output

Command Expected Input Explanation
--outFileCorMatrix FILENAME Output file name for the correlation matrix (tabulated text file). (default: None)
--outRawCounts FILENAME Output file name to save the bin counts (tabulated text file). (default: None)
--plotFileFormat {png, emf, eps, pdf, svg} If given, this option overrides the image format based on the ending given in --plotFile. (default: None)

bamFingerprint

Mandatory arguments

Command Expected Input Explanation
--bamfiles FILENAME List of sorted BAM files (default: None)
--labels LIST List of labels to use in the output. If not given the file names will be used instead. Separate the labels by space. (default: None)
--binSize INT Length in base pairs for a window used to sample the genome. (default: 500)
--fragmentLength INT Length of the average fragment size. (default: 200)
--numberOfSamples INT Number of bins, sampled from the genome to compute the average number of reads. (default: 500000)
--skipZeros If set, then zero counts that happen for all BAM files given are ignored. This will result in a reduced number of read counts than the the specified in --numberOfSamples (default: False)

bamFingerprint: optional arguments

Command Expected Input Explanation
--region CHR:START:END Region of the genome to limit the operation to - this is useful when testing parameters to reduce the computing time. The format is chr:start:end, for example --region chr10 or --region chr10:456700:891000. (default: None)
--numberOfProcessors INT Number of processors to use. Type "max/2" to use half the maximum number of processors or "max" to use all available processors. (default: max/2)
--verbose Set to see processing messages. (default: False)

bamFingerprint: Processing options

Command Expected Input Explanation
--doNotExtendPairedEnds If set, reads are not extended to match the fragment length reported in the BAM file, instead they will be extended to match the --fragmentLength. Default is to extend the reads if paired end information is available. (default: False)
--ignoreDuplicates If set, reads that have the same orientation and start position will be considered only once. If reads are paired, the mate position also has to coincide to ignore a read. (default: False)
--minMappingQuality INT If selected and accompanied by a number, only reads that have a mapping quality score higher than the given number are considered (e.g. --minMappingQuality 10)(default: None)

bamFingerprint: output options

Command Expected Input Explanation
--plotFile FILENAME File name to save the image file containing a plot of the fingerprint, for example MyPlot.png (default: None)
--outRawCounts FILENAME Output file name to save the bin counts (default: None)
--plotFileFormat {png, emf, eps, pdf, svg} image format type. If given, this option overrides the image format based on the plotFile ending. (default: None)

bamCoverage

Mandatory arguments

Command Expected Input Explanation
--bam FILENAME BAM file to process (default: None)
--bamIndex FILENAME Index for the BAM file. Default is to consider the path of the BAM file adding the .bai suffix. (default: None)
--scaleFactor NUMBER Indicate a number that you would like to use as a fixed scaling factor instead of the scaling factor calculated by the --normalizeUsingSequencingDepth option. (default: 1)
--normalizeTo1x NUMBER Report read coverage normalized to 1x sequencing depth (also known as Reads Per Genomic Content (RPGC)). Sequencing depth is defined as: (total number of mapped reads * fragment length) / effective genome size. To use this option, the effective genome size has to be indicated after the command**. The effective genome size is the portion of the genome that is mappable. Large fractions of the genome are stretches of NNNN that should be discarded. Also, if repetitive regions were not included in the mapping of reads, the effective genome size needs to be adjusted accordingly. Common values are: mouse/mm9: 2150570000, human/hg19:2451960000, D.melanogaster/dm3:121400000 and C.elegans/ce10:93260000. See Table 2 of http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030377 or http://www.nature.com/nbt/journal/v27/n1/fig_tab/nbt.1518_T1.html for several effective genome sizes. (default: None)
--normalizeUsingRPKM Use Reads Per Kilobase per Million reads to normalize the number of reads per bin. The formula is: RPKM (per bin) = number of reads per bin / ( number of mapped reads ( in millions) * bin length (kb) ) (default: False)
--ignoreForNormalization LIST A list of chromosome names separated by comma and limited by quotes, containing those chromosomes that want to be excluded for computing the normalization. For example, --ignoreForNormalization "chrX, chrM" (default: None)

bamCoverage: optional arguments

Command Expected Input Explanation
--binSize INT Size of the bins in bp for the output of the bigWig/bedGraph file. (default: 50)
--region CHR:START:END Region of the genome to limit the operation to - this is useful when testing parameters to reduce the computing time. The format is chr:start:end, for example --region chr10 or --region chr10:456700:891000. (default: None)
--numberOfProcessors INT Number of processors to use. Type "max/2" to use half the maximum number of processors or "max" to use all available processors. (default: max/2)
--verbose Set to see processing messages. (default: False)

bamCoverage: BAM to bedGraph/bigWig processing options

Command Expected Input Explanation
--fragmentLength INT Length of the average fragment size. Reads will be extended to match this length unless they are paired- end, in which case they will be extended to match the fragment length. If this value is set to the read length or smaller, the read will not be extended. Warning the fragment length affects the normalization to 1x (see --normalizeUsingSequencingDepth). The formula to normalize using the sequencing depth is genomeSize/(number of mapped reads * fragmentLength). NOTE: If the BAM files contain mated and unmated paired-end reads, unmated reads will be extended to match the --fragmentLength. (default: 200)
--smoothLength INT The smooth length defines a window, larger than the binSize, to average the number of reads. For example, if the --binSize is set to 20 bp and the --smoothLength is set to 60 bp, then, for each binSize the average of it and its left and right neighbors is considered. Any value smaller than the --binSize will be ignored and no smoothing will be applied. (default: None)
--doNotExtendPairedEnds If set, reads are not extended to match the fragment length reported in the BAM file, instead they will be extended to match the --fragmentLength. Default is to extend the reads if paired end information is available. (default: False)
--ignoreDuplicates If set, reads that have the same orientation and start position will be considered only once. If reads are paired, the mate position also has to coincide to ignore a read. (default: False)
--minMappingQuality INT If set, only reads that have a mapping quality score higher than --minMappingQuality are considered. (default: None)

bamCoverage: output options

Command Expected Input Explanation
--outFileName FILENAME Output file name. (default: None)
--outFileFormat {bigwig,bedgraph} Output file type.

bamCompare

Mandatory arguments

Command Expected Input Explanation
--bamfile1 FILENAME Sorted BAM file 1. Usually the BAM file for the treatment. (default: None)
--bamIndex1 FILENAME Index for the BAM file 1. Default is to consider the path of the BAM file and adding the .bai suffix. (default: None)
--bamfile2 FILENAME Sorted BAM file 2. Usually the BAM file for the control. (default: None)
--bamIndex2 FILENAME Index for the BAM file 1. Default is to consider the path of the BAM file adding the .bai suffix. (default: None)
--scaleFactorsMethod {readCount,SES} Method for scaling the samples. SES is only recommended when signal and noise are well separated which can be seen in the bamFingerprint plot. (default: readCount)
--sampleLength INT Length in base pairs used to sample the genome and compute the size or scaling factors to compare the two BAM files (default: 1000)
--scaleFactors NUMBER:NUMBER Set this parameter to avoid the computation of scaleFactors. The format is scaleFactor1:scaleFactor2. For example 0.7:1 to scale the first BAM file by 0.7 while not scaling the second BAM file (default: None)
--pseudocount NUMBER small number to avoid log2(x/0) (default: 1)
--ratio {log2, ratio, subtract, add, reciprocal_ratio} The default is to output the log2ratio between the two samples. The reciprocal ratio returns the the negative of the inverse of the ratio if the ratio is less than 0. The resulting values are interpreted as negative fold changes. (default: log2)
--normalizeUsingRPKM only to be set when --ratio subtract is selected If you would like to get the difference between 2 BAM files (and hence set --ratio subtract), the final score will be normalized for sequencing depth so that the file can be compared to other samples from different sequencing runs, too. The default normalization method is R Per Kilobase per Million reads. The formula is: RPKM (per bin) = #reads per bin / ( # of mapped reads (millions) * bin length (KB)). Example usage: --normalizeUsingRPKM
--normalizeTo1x NUMBER (only when --ratio subtract) The difference will be reported normalized for sequencing depth. The default method is RPKM (see above). If you would like to report the normalized coverage to 1x sequencing depth, set this option and indicate the effective genome size. Sequencing depth is defined as the total number of mapped reads*fragment length / effective genome size. To use this option, the effective genome size has to be given. Common values are: mouse/mm9: 2150570000, human/hg19:2451960000, D.melanogaster/dm3:121400000 and C.elegans/ce10:93260000. Example usage: --normalizeTo1x 2150570000
--numberOfSamples INT How many times the genome should be sampled to compute the scaling factors (default: 100000)
--missingDataAsZero {yes,no} This parameter determines if missing data should be treated as zeros. If set to "no", missing data will be ignored and not included in the output file. Missing data is defined as those regions for which both BAM files have 0 reads. (default: yes)
--ignoreForNormalization LIST A list of chromosome names separated by comma and limited by quotes, containing those chromosomes that you want to be excluded for computing the normalization. For example, --ignoreForNormalization "chrX, chrM" (default: None)

bamCompare: optional arguments

Command Expected Input Explanation
--version show program's version number and exit
--binSize INT Size of the bins in bp for the output of the bigWig/bedGraph file. (default: 50)
--region CHR:START:END Region of the genome to limit the operation to - this is useful when testing parameters to reduce the computing time. The format is chr:start:end, for example --region chr10 or --region chr10:456700:891000. (default: None)
--numberOfProcessors INT Number of processors to use. Type "max/2" to use half the maximum number of processors or "max" to use all available processors. (default: max/2)
--verbose Set to see processing messages.

bamCompare: BAM to bedGraph/bigWig processing options

Command Expected Input Explanation
--fragmentLength INT Length of the average fragment size. Reads will be extended to match this length unless they are paired- end, in which case they will be extended to match the fragment length. If this value is set to the [read][] length or smaller, the read will not be extended. Warning the fragment length affects the normalization to 1x (see --normalizeUsingSequencingDepth). The formula to normalize using the sequencing depth is genomeSize/(number of mapped reads * fragmentLength). NOTE: If the BAM files contain mated and unmated paired-end reads, unmated reads will be extended to match the --fragmentLength. (default: 200)
--smoothLength INT The smooth length defines a window, larger than the binSize, to average the number of reads. For example, if the --binSize is set to 20 bp and the --smoothLength is set to 60 bp, then, for each binSize the average of it and its left and right neighbors is considered. Any value smaller than the --binSize will be ignored and no smoothing will be applied. (default: None)
--doNotExtendPairedEnds If set, reads are not extended to match the fragment length reported in the BAM file, instead they will be extended to match the --fragmentLength. Default is to extend the reads if paired end information is available. (default: False)
--ignoreDuplicates If set, reads that have the same orientation and start position will be considered only once. If reads are paired, the mate position also has to coincide to ignore a read. (default: False)
--minMappingQuality INT If selected and accompanied by a number, only reads that have a mapping quality score higher than the given number are considered (e.g. --minMappingQuality 10)(default: None)

bamCompare: output options

Command Expected Input Explanation
--outFileName FILENAME Output file name. (default: None)
--outFileFormat {bigwig, bedgraph} Output file type.


[read]: https://github.com/fidelram/deepTools/wiki/Glossary#terminology "the DNA piece that was actually sequenced ("read") by the sequencing machine (usually between 30 to 100 bp long, depending on the read-length of the sequencing protocol)" [input]: https://github.com/fidelram/deepTools/wiki/Glossary#terminology "confusing, albeit commonly used name for the 'no-antibody' control sample for ChIP experiments"

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