The purpose of this program is to generate metrics at single nucleotide positions.
There are number of metrics generated which can be useful for filtering out false positive calls.
Help is currently available on the commandline if you do not supply the program
any arguments.
Usage: bam-readcount <bam_file> [region]
Available options:
-h [ --help ] produce this message
-q [ --min-mapping-quality ] arg (=0) minimum mapping quality of reads used
for counting.
-b [ --min-base-quality ] arg (=0) minimum base quality at a position to
use the read for counting.
-d [ --max-count ] arg (=10000000) max depth to avoid excessive memory
usage.
-l [ --site-list ] arg file containing a list of regions to
report readcounts within.
-f [ --reference-fasta ] arg reference sequence in the fasta format.
-D [ --print-individual-mapq ] arg report the mapping qualities as a comma
separated list.
-p [ --per-library ] report results by library.
-w [ --max-warnings ] arg maximum number of warnings of each type
to emit. -1 gives an unlimited number.
-i [ --insertion-centric ] generate indel centric readcounts.
Reads containing insertions will not be
included in per-base countsThis program reports readcounts for each base at each position requested.
It also reports the average base quality of these bases and mapping qualities of
the reads containing each base.
The list of regions should be formatted as chromosome start and end. Each field should be tab separated and coordinates should be 1-based.
The optional region specification on the command line should follow the same format as that used for samtools (chr:start-stop)
The output format is as follows:
chr position reference_base depth base:count:avg_mapping_quality:avg_basequality:avg_se_mapping_quality:num_plus_strand:num_minus_strand:avg_pos_as_fraction:avg_num_mismatches_as_fraction:avg_sum_mismatch_qualities:num_q2_containing_reads:avg_distance_to_q2_start_in_q2_reads:avg_clipped_length:avg_distance_to_effective_3p_end ...Each tab separated field contains the following information, each one is tab-separated:
base → the base that all reads following in this field contain at the reported position i.e. C
count → the number of reads containing the base
avg_mapping_quality → the mean mapping quality of reads containing the base
avg_basequality → the mean base quality for these reads
avg_se_mapping_quality → mean single ended mapping quality
num_plus_strand → number of reads on the plus/forward strand
num_minus_strand → number of reads on the minus/reverse strand
avg_pos_as_fraction → average position on the read as a fraction (calculated with respect to the length after clipping)
avg_num_mismatches_as_fraction → average number of mismatches on these reads per base
avg_sum_mismatch_qualities → average sum of the base qualities of mismatches in the reads
num_q2_containing_reads → number of reads with q2 runs at the 3’ end
avg_distance_to_q2_start_in_q2_reads → average distance of position (as fraction of unclipped read length) to the start of the q2 run
avg_clipped_length → average clipped read length of reads
avg_distance_to_effective_3p_end → average distance to the 3’ prime end of the read (as fraction of unclipped read length)
Beginning with version 0.5.0, bam-readcount can now output counts per each library. Libraries are reported with the same metrics as in
normal counting, but each library is listed by name and its metrics denoted by curly braces as follows:
chr position reference_base depth libname { base:count:avg_mapping_quality:avg_basequality:avg_se_mapping_quality:num_plus_strand:num_minus_strand:avg_pos_as_fraction:avg_num_mismatches_as_fraction:avg_sum_mismatch_qualities:num_q2_containing_reads:avg_distance_to_q2_start_in_q2_reads:avg_clipped_length:avg_distance_to_effective_3p_end } ...- git
- cmake 2.8+ (cmake.org)
- For APT-based systems (Debian, Ubuntu), install the following packages:
sudo apt-get install build-essential git-core cmake zlib1g-dev libncurses-dev patch- For RPM-based systems (Fedora, CentOS, RHEL), install the following packages instead:
sudo yum groupinstall "Development tools"
sudo yum install zlib-devel ncurses-devel cmake patchNote that for some RPM based systems (like RHEL), you will need to install cmake 2.8 or greater yourself as the packaged version is much older.
- Recursively clone the git repository
git clone https://github.com/genome/bam-readcount.git- bam-readcount does not support in source builds. Create a new build directory, enter it, and run:
cmake /path/to/bam-readcount/repo
make deps
makeThe binary can then be found in the bin/ subdirectory of your build directory.
These errors should no longer occur as of commit e48b05cbb9a9222d7585958f9704b147f0a4cbea. For earlier commits, bam-readcount contains a git submodule called build-common. It contains helper modules for cmake. If you downloaded the source as a tarball from github or forgot to do a recursive clone using git, then you will not have this submodule and will see cmake errors. If you are using git, we recommend you go back and use the —recursive option when cloning the repository. If you cannot use git, follow the instructions below to remedy the situation.
- Download the build-common module separately here
- Extract that tarball, and rename the directory it creates to ‘build-common’.
- Replace the empty build-common subdirectory in the sniper directory with directory you just created.
- Resume following the build instructions.
This is a known bug that will be addressed in future versions. In most cases, you will only need read counts on a subset of positions and specifying the sites of interest is functional. Should you need to run the entire genome through then running by chromosome and concatenating the results should be a decent workaround (e.g. like bam-readcount -f ref.fasta some.bam chr1).
Please first search Biostar and then ask a question there if needed. We automatically monitor Biostar for questions related to our tools.