A highly parallelized utility for analyzing metrics at a per-base level.
If a metric is missing, or performance is lacking. Please file a bug/feature issue. This tool aims to improve upon bam-readcount.
```bash conda install -c bioconda -c conda-forge perbase
cargo install perbase ```
* Check version, conda lags behind what you will find in the release page
The simple-depth tool walks over every position in the BAM/CRAM file and calculates the depth, as well as the number of each nucleotide at the given position. Additionally, it counts the numbers of Ins/Dels at each position.
The output columns are as follows:
| Column | Description | | -------- | -------------------------------------------------------------------------------------------------- | | REF | The reference sequence name | | POS | The 1-based position on the reference sequence | | DEPTH | The total depth at the position SUM(A, C, T, G, DEL) | | A | Total A nucleotides seen at this position | | C | Total C nucleotides seen at this position | | G | Total G nucleotides seen at this position | | T | Total T nucleotides seen at this position | | N | Total N nucleotides seen at this position | | INS | Total insertions that start at the base to the right of this position | | DEL | Total deletions covering this position | | REF_SKIP | Total reference skip operations covering this position | | FAIL | Total reads failing filters that covered this position (their bases were not counted toward depth) |
bash
perbase simple-depth ./test/test.bam
Example output
text
REF POS DEPTH A C G T N INS DEL FAIL REF_SKIP
chr2 1 1 1 0 0 0 0 0 0 0 0
chr2 2 1 1 0 0 0 0 1 0 0 0
chr2 3 1 0 0 1 0 0 0 0 0 0
chr2 4 1 1 0 0 0 0 0 0 0 0
chr2 5 2 2 0 0 0 0 0 0 0 0
chr2 6 2 2 0 0 0 0 0 0 0 0
chr2 7 2 1 0 0 0 0 0 1 0 0
chr2 8 2 1 0 0 0 0 0 1 0 0
chr2 9 2 1 0 0 0 0 0 1 0 0
chr2 10 3 2 0 0 0 0 0 1 0 0
chr2 11 3 2 0 0 0 0 0 1 0 0
chr2 12 3 3 0 0 0 0 0 0 0 0
chr2 13 3 2 0 0 0 0 0 0 0 1
chr2 14 3 2 0 0 0 0 0 0 0 1
chr2 15 4 3 0 0 0 0 0 0 0 1
chr2 16 4 2 0 0 1 0 0 0 0 1
chr2 17 4 3 0 0 0 0 0 0 0 1
If the --mate-fix flag is passed, each position will first check if there are any mate overlaps and choose the mate with the hightest MAPQ, breaking ties by choosing the first mate that passes filters. Mates that are discarded are not counted toward FAIL or DEPTH.
The output can be compressed and indexed as follows:
```bash perbase simple-depth ./test/test.bam | bgzip > output.tsv.gz tabix -S 1 -s 1 -b 2 -e 2 ./output.tsv.gz
tabix output.tsv.gz chr1:5-10 ```
Usage:
```text
Usage: perbase simple-depth
Calculate the depth at each base, per-nucleotide. Takes an indexed BAM/CRAM as
Options: -r, --ref-fasta indexed reference fasta, set if using CRAM -b, --bed-file a BED file containing regions of interest. If specified, only bases from the given regions will be reported on -o, --output output path. DEFAULT: stdout -t, --threads the number of threads to use. DEFAULT: max_available -c, --chunksize the ideal number of basepairs each worker receives. Total bp in memory at one time is (threads - 2) * chunksize -f, --include-flags SAM flags to include. DEFAULT: 0 -F, --exclude-flags SAM flags to exclude, recommended 3848. DEFAULT: 0 -m, --mate-fix fix overlapping mates counts, see docs for full details. DEAFAULT: off -q, --min-mapq minimum mapq for a read to count toward depth. DEFAULT: 0 --help display usage information ```