Awesome
Synopsis
Get the readcounts at a locus by piping samtools mpileup output. The mpileup can contain one or several samples. This program has been tested on samtools v1.3.1
Install samtools
Compile mpileup2readcounts :
g++ -std=c++11 -O3 mpileup2readcounts.cc -o mpileup2readcounts
Usage
samtools mpileup -f ref.fa -l regions.bed BAM/*.bam | sed 's/ / * */g' | ./mpileup2readcounts 0 -5 false 3 0
Samtools arguments :
- FASTA file
- bed file
- BAM files : several samples can be parsed
Four options for mpileup2readcounts :
- 0 to parse all sample otherwise specify the number of the sample (for example 1 for the first sample)
- BQcut : base quality score cutoff for each mapped/unmapped base, only those larger than cutoff will be output in the result, to use no filter set BQcut to -5
- true to ignore indels
- min_ao : minimum number of non-ref reads in at least one sample to consider a site
- min_af : minimum allelic fraction in at least one sample to consider a site
Example output
| chr | loc | ref | depth | A | T | C | G | a | t | c | g | Insertion | Deletion | depth | A | T | C | G | a | t | c | g | Insertion | Deletion |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 17 | 7572814 | C | 28 | 0 | 3 | 23 | 0 | 0 | 0 | 2 | 0 | NA | NA | 8 | 0 | 0 | 8 | 0 | 0 | 0 | 0 | 0 | NA | NA |
| 17 | 7572817 | C | 32 | 2 | 0 | 26 | 0 | 2 | 0 | 2 | 0 | NA | NA | 8 | 0 | 0 | 8 | 0 | 0 | 0 | 0 | 0 | NA | NA |
| 17 | 7579643 | C | 48 | 0 | 0 | 9 | 0 | 0 | 0 | 39 | 0 | NA | 4:ccccagccctccaggt|2:CCCCAGCCCTCCAGGT | 9 | 0 | 0 | 6 | 0 | 0 | 0 | 3 | 0 | NA | NA |
Line content
Common information for all samples:
- chromosome
- position on the chromosome
- reference base
For each sample :
- depth
- ATCG/atcg count
- insertions
- deletions : in the example, for the first sample 6 deletions starting from position 7579643 + 1 are found
Using GNU parallel tool
We first remove overlap in the bed using bedtools. Note that the header is removed here:
grep -v '^track' regions.bed | sort -k1,1 -k2,2n | bedtools merge -i stdin | awk '{print $1"\t"$2"\t"$3}' | sed 's/[[:space:]]/:/' | sed 's/[[:space:]]/-/' | parallel --keep-order "samtools mpileup -f ref.fa --region {} test.bam | sed 's/ / * */g' | mpileup2readcounts 0 -5 false 0 | tail -n +2"