ANGSD: Analysis of next generation Sequencing Data

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2d SFS Estimation: Difference between revisions

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* Assume you have a 14 bamfiles for population in the file '''pop2.list'''
* Assume you have a 14 bamfiles for population in the file '''pop2.list'''
* Assume you have a fastafile containing the ancestral state in the '''anc.fa'''
* Assume you have a fastafile containing the ancestral state in the '''anc.fa'''
* Assume we are only interested in '''chr1'''


Let's start by finding the positions for which we have data in population1 and population2
Let's start by finding the positions for which we have data in population1 and population2
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If we were interested in estimating the 1d sfs for each population we could do it like this using the [[realSFS]] program. (See more on [[SFS Estimation |page]] )
If we were interested in estimating the 1d sfs for each population we could do it like this using the [[realSFS]] program. (See more on [[SFS Estimation |page]] )
<pre>
<pre>
realSFS pop1.saf 24 -P 24 >pop1.saf.sfs
#sfs for pop1
realSFS pop2.saf 28 -P 24 >pop2.saf.sfs
realSFS pop1.saf.idx -P 24 >pop1.saf.sfs
#first argument is saf file, second argument is the number of chromosomes, -P 24 is the number of cores we want to use
#sfs for pop2
</pre>
realSFS pop2.saf.idx -P 24 >pop2.saf.sfs
Now we find the positions that occurs both in population1 and population2 using the '''uniq''' POSIX program.
#2d sfs for pop1 and pop2
<pre>
#first argument is saf file, -P 24 is the number of cores we want to use
gunzip -c pop1.saf.pos.gz pop2.saf.pos.gz|sort  -S 50%|uniq -d|sort -k1,1  -S 50% >intersect.txt
realSFS pop1.saf.idx pop2.saf.idx -P 24
</pre>
And now we redo the angsd sample allelefrequence calculation by conditioning on the sites that occur in both populations
 
<pre>
# as always you can add -minMapQ 1 and -minQ 20 to only keep high quality data.
angsd -GL 1 -b pop1.list -anc anc.fa -r chr1: -P 10 -out pop1 -sites intersect.txt -doSaf 1
angsd -GL 1 -b pop2.list -anc anc.fa -r chr1: -P 10 -out pop2 -sites intersect.txt -doSaf 1
</pre>
Notice that the last 2 commands will overwrite the: '''pop1.saf,pop1.saf.pos''' and '''pop2.saf,pop2.saf.pos''' files.
 
 
And we now estimate the joint site frequency spectra by using the [[realSFS]] program
And we now estimate the joint site frequency spectra by using the [[realSFS]] program
<pre>
<pre>
realSFS 2dsfs pop1.saf pop2.saf  24 28 -P 24 >2dsfs.sfs
</pre>
The output is then located in a nice matrix format(25x29) in the file: '''2dsfs.sfs'''. Good luck visualising it, some people are using dadi, we have been using heat maps in R.
The output is then located in a nice matrix format(25x29) in the file: '''2dsfs.sfs'''. Good luck visualising it, some people are using dadi, we have been using heat maps in R.

Revision as of 14:37, 11 May 2015

Angsd can estimate a 2d site frequency spectrum. This is an extension of the 1d site frequency spectrum method. Never versions of ANGSD can estimate even higher dimensions (upto 4)

And is best explained by a full example.

Example

  • Assume you have a 12 bamfiles for population in the file pop1.list
  • Assume you have a 14 bamfiles for population in the file pop2.list
  • Assume you have a fastafile containing the ancestral state in the anc.fa

Let's start by finding the positions for which we have data in population1 and population2

# as always you can add -minMapQ 1 and -minQ 20 to only keep high quality data.
angsd -GL 1 -b pop1.list -anc anc.fa -r chr1: -P 10 -out pop1 -doSaf 1
angsd -GL 1 -b pop2.list -anc anc.fa -r chr1: -P 10 -out pop2 -doSaf 1

Each run will generate 2 files of interest: pop1.saf,pop1.saf.pos and pop2.saf,pop2.saf.pos

If we were interested in estimating the 1d sfs for each population we could do it like this using the realSFS program. (See more on page )

#sfs for pop1
realSFS pop1.saf.idx -P 24 >pop1.saf.sfs
#sfs for pop2
realSFS pop2.saf.idx -P 24 >pop2.saf.sfs
#2d sfs for pop1 and pop2
#first argument is saf file,  -P 24 is the number of cores we want to use
realSFS pop1.saf.idx pop2.saf.idx -P 24
And we now estimate the joint site frequency spectra by using the realSFS program
The output is then located in a nice matrix format(25x29) in the file: 2dsfs.sfs. Good luck visualising it, some people are using dadi, we have been using heat maps in R.