ANGSD: Analysis of next generation Sequencing Data
Latest tar.gz version is (0.938/0.939 on github), see Change_log for changes, and download it here.
Fasta
This option creates a fasta.gz file from a sequencing data file (BAM file). The function uses genome information in the BAM header to determine the length and chromosome names. For the sites without data an "N" is written.
<classdiagram type="dir:LR">
[Single BAM file{bg:orange}]->[Sequence data|Random base (-doFasta 1);Consensus base (-doFasta 2);Highest EBD (-doFasta 3)]
[sequence data]->doFasta[fasta file{bg:blue}]
</classdiagram>
<classdiagram type="dir:LR">
[Multiple BAM files{bg:orange}]->[Sequence data|Random base (-doFasta 1);Consensus base (-doFasta 2)]
[sequence data]->doFasta[fasta file{bg:blue}]
</classdiagram>
This can be used as input for the ANGSD analysis:
Brief Overview
./angsd -doFasta -> Wed Nov 16 16:58:52 2016 -------------- abcWriteFasta.cpp: -doFasta 0 1: use a random (non N) base (needs -doCounts 1) 2: use the most common (non N) base (needs -doCounts 1) 3: use the base with highest ebd (under development) -basesPerLine 50 (Number of bases perline in output file) -explode 0 print chromosome where we have no data (0:no,1:yes) -rmTrans 0 remove transitions as different from -ref bases (0:no,1:yes) -ref (null) reference fasta, only used with -rmTrans 1 -seed 0 use non random seed of value 1
This function will dump a fasta file, the full header information from the SAM/BAM file will be used. This means that a fasta will be generated for the entire chromosome even if '-r/-rf -sites' is used.
Options
- -doFasta 1
- sample a random base at each position. N's or filtered based are ignored. The "-doCounts 1" options for allele counts is needed in order to determine the most common base. If multiple individuals are used the four bases are counted across individuals.
- -doFasta 2
- use the most common base. In the case of ties a random base is chosen among the bases with the same maximum counts. N's or filtered based are ignored. The "-doCounts 1" options for allele counts is needed in order to determine the most common base. If multiple individuals are used the four bases are counted across individuals.
- -doFasta 3
- use the base with thie highest effective depth (EBD). This only works for one individual
- -basesPerLine [INT]
Number of bases perline in output fasta file (default is 50)
- -explode [INT]
0 (default) only output chromosomes with data. 1: write out all chromosomes
- -rmTrans [INT]
0 (default) all sites are used. 1: Remove transition. Here transitions are determined using a fasta file such as a reference genome.
- -ref [fileName]
a fasta file used to determine if a site is a transitions (needed when using -rmTrans 1 is used)
- -seed [INT]
Use a seed in order to replicate results
For filters see Filters
Output
Output is a fasta file, a normal looking fast file. Nothing special about this. For -doFasta 1, sometimes its big letters sometime small letters. This is due to the results being copied directly from the sequencing data. So small/big letters correspond to which strand for the original data. For the consensus fasta all letters are capital letters.
Example
Create a fasta file bases from a random samples of bases.
./angsd -i bams/smallNA07056.mapped.ILLUMINA.bwa.CEU.low_coverage.20111114.bam -doFasta 1
EBD
For four bases we have 4 different EBD, each EBD is the product of the mapping quality and scores for the base under consideration. The EBD is the effective base depth, as defined by [1]:
where is a certain base, is the number of reads with base