Bio::PopGen Statistics
SummaryIncluded librariesPackage variablesSynopsisDescriptionGeneral documentationMethods
Summary
Bio::PopGen::Statistics - Population Genetics statistical tests
Package variables
No package variables defined.
Included modules
Bio::Root::Root
Inherit
Bio::Root::Root
Synopsis
  use Bio::PopGen::Statistics;
  use Bio::AlignIO;
  use Bio::PopGen::IO;
  use Bio::PopGen::Simulation::Coalescent;

  my $sim = new Bio::PopGen::Simulation::Coalescent( -sample_size => 12);

  my $tree = $sim->next_tree;

  $factory->add_Mutations($tree,20);

  my $stats = new Bio::PopGen::Statistics();
  my $individuals = [ $tree->get_leaf_nodes];
  my $pi = $stats->pi($individuals);
  my $D  = $stats->tajima_d($individuals);

  # Alternatively to do this on input data from
  # See the tests in t/PopGen.t for more examples
  my $parser = new Bio::PopGen::IO(-format => 'prettybase',
                                   -file   => 't/data/popstats.prettybase');
  my $pop = $parser->next_population;
  # Note that you can also call the stats as a class method if you like
  # the only reason to instantiate it (as above) is if you want
  # to set the verbosity for debugging
  $pi     = Bio::PopGen::Statistics->pi($pop);   
  $theta  = Bio::PopGen::Statistics->theta($pop);

  # Pi and Theta also take additional arguments,
  # see the documentation for more information  

  # To come -- examples for creating pops/individuals from
  # Aligned sequence data
Description
This object is intended to provide implementations some standard
population genetics statistics about alleles in populations.
This module was previously named Bio::Tree::Statistics.
This object is a place to accumulate routines for calculating various
statistics from the coalescent simulation, marker/allele, or from
aligned sequence data given that you can calculate alleles, number of
segregating sites.
Currently implemented:
Fu and Li's D (fu_and_li_D)
Fu and Li's D* (fu_and_li_D_star)
Fu and Li's F (fu_and_li_F)
Tajima's D (tajima_D)
theta (theta)
pi (pi) - number of pairwise differences
composite_LD (composite_LD)
In all cases where a the method expects an arrayref of
Bio::PopGen::IndividualI objects and Bio::PopGen::PopulationI
object will also work. Fu Y.X and Li W.H. (1993) "Statistical Tests of Neutrality of
Mutations." Genetics 133:693-709.
Fu Y.X. (1996) "New Statistical Tests of Neutrality for DNA samples
from a Population." Genetics 143:557-570.
Tajima F. (1989) "Statistical method for testing the neutral mutation
hypothesis by DNA polymorphism." Genetics 123:585-595.
Methods
fu_and_li_DDescriptionCode
fu_and_li_D_starDescriptionCode
fu_and_li_FDescriptionCode
fu_and_li_F_starDescriptionCode
tajima_DDescriptionCode
piDescriptionCode
thetaDescriptionCode
singleton_countDescriptionCode
segregating_sites_countDescriptionCode
heterozygosityDescriptionCode
derived_mutationsDescriptionCode
composite_LDDescriptionCode
Methods description
fu_and_li_Dcode    nextTop
 Title   : fu_and_li_D
 Usage   : my $D = $statistics->fu_an_li_D(\@ingroup,$extmutations);
 Function: Fu and Li D statistic for a list of individuals
           given an outgroup and the number of external mutations
           (either provided or calculated from list of outgroup individuals)
 Returns : decimal
 Args    : $individuals - array refence which contains ingroup individuals 
           (Bio::PopGen::Individual or derived classes)
$extmutations - number of external mutations OR
arrayref of outgroup individuals
fu_and_li_D_starcodeprevnextTop
 Title   : fu_and_li_D_star
 Usage   : my $D = $statistics->fu_an_li_D_star(\@individuals);
 Function: Fu and Li's D* statistic for a set of samples
            Without an outgroup
 Returns : decimal number
 Args    : array ref of Bio::PopGen::IndividualI objects
OR
Bio::PopGen::PopulationI object
fu_and_li_FcodeprevnextTop
 Title   : fu_and_li_F
 Usage   : my $D = Bio::PopGen::Statistics->fu_and_li_F(\@ingroup,$ext_muts);
 Function: Calculate Fu and Li's F on an ingroup with either the set of 
           outgroup individuals, or the number of external mutations
 Returns : decimal number
 Args    : array ref of Bio::PopGen::IndividualI objects for the ingroup
OR a Bio::PopGen::PopulationI object
number of external mutations OR list of individuals for the outgroup
fu_and_li_F_starcodeprevnextTop
 Title   : fu_and_li_F_star
 Usage   : my $D = Bio::PopGen::Statistics->fu_and_li_F_star(\@ingroup);
 Function: Calculate Fu and Li's F* on an ingroup without an outgroup
           It uses count of singleton alleles instead 
 Returns : decimal number
 Args    : array ref of Bio::PopGen::IndividualI objects for the ingroup
OR
Bio::PopGen::PopulationI object
tajima_DcodeprevnextTop
 Title   : tajima_D
 Usage   : my $D = Bio::PopGen::Statistics->tajima_D(\@samples);
 Function: Calculate Tajima's D on a set of samples 
 Returns : decimal number
 Args    : array ref of Bio::PopGen::IndividualI objects
OR
Bio::PopGen::PopulationI object
picodeprevnextTop
 Title   : pi
 Usage   : my $pi = Bio::PopGen::Statistics->pi(\@inds)
 Function: Calculate pi (...explain here...) given a list of individuals 
           which have the same number of markers/sites/mutation as 
           available from the get_Genotypes() call in 
           Bio::PopGen::IndividualI
Returns : decimal number
Args : Arg1= array ref of Bio::PopGen::IndividualI objects
which have markers/mutations. We expect all individuals to
have a marker - we will deal with missing data as a special case.
OR
Arg1= Bio::PopGen::PopulationI object. In the event that
only allele frequency data is available, storing it in
Population object will make this available.
num sites [optional], an optional second argument (integer)
which is the number of sites, then pi returned is pi/site.
thetacodeprevnextTop
 Title   : theta
 Usage   : my $theta = Bio::PopGen::Statistics->theta($sampsize,$segsites);
 Function: Calculates theta (...explanation here... ) from the sample size 
           and the number of segregating sites.
           Providing the third parameter, total number of sites will
           return theta per site          
 Returns : decimal number 
 Args    : sample size (integer),
           num segregating sites (integer)
           total sites (integer) [optional] (to calculate theta per site)
           OR
           provide an arrayref of the Bio::PopGen::IndividualI objects
total sites (integer) [optional] (to calculate theta per site)
OR
provide an Bio::PopGen::PopulationI object
total sites (integer)[optional]
singleton_countcodeprevnextTop
 Title   : singleton_count
 Usage   : my ($singletons) = Bio::PopGen::Statistics->singleton_count(\@inds)
 Function: Calculate the number of mutations/alleles which only occur once in
           a list of individuals for all sites/markers
 Returns : (integer) number of alleles which only occur once (integer)
 Args    : arrayref of Bio::PopGen::IndividualI objects
OR
Bio::PopGen::PopulationI object
segregating_sites_countcodeprevnextTop
 Title   : segregating_sites_count
 Usage   : my $segsites = Bio::PopGen::Statistics->segregating_sites_count
 Function: Gets the number of segregating sites (number of polymorphic sites)
 Returns : (integer) number of segregating sites
 Args    : arrayref of Bio::PopGen::IndividualI objects 
OR
Bio::PopGen::PopulationI object
heterozygositycodeprevnextTop
 Title   : heterozygosity
 Usage   : my $het = Bio::PopGen::Statistics->heterozygosity($sampsize,$freq1);
 Function: Calculate the heterozgosity for a sample set for a set of alleles
 Returns : decimal number
 Args    : sample size (integer)
           frequency of one allele (fraction - must be less than 1)
           [optional] frequency of another allele - this is only needed
                      in a non-binary allele system
Note : p^2 + 2pq + q^2
derived_mutationscodeprevnextTop
 Title   : derived_mutations
 Usage   : my $ext = Bio::PopGen::Statistics->derived_mutations($ingroup,$outgroup);
 Function: Calculate the number of alleles or (mutations) which are ancestral
           and the number which are derived (occurred only on the tips)
 Returns : array of 2 items - number of external and internal derived 
           mutation
 Args    : ingroup - Bio::PopGen::IndividualIs arrayref OR 
Bio::PopGen::PopulationI
outgroup- Bio::PopGen::IndividualIs arrayref OR
Bio::PopGen::PopulationI OR
a single Bio::PopGen::IndividualI
composite_LDcodeprevnextTop
 Title   : composite_LD
 Usage   : %matrix = Bio::PopGen::Statistics->composite_LD($population);
 Function: Calculate the Linkage Disequilibrium 
           This is for calculating LD for unphased data. 
           Other methods will be appropriate for phased haplotype data.

 Returns : Hash of Hashes - first key is site 1,second key is site 2
           and value is LD for those two sites.
           my $LDarrayref = $matrix{$site1}->{$site2};
           my ($ldval, $chisquared) = @$LDarrayref;
 Args    : Bio::PopGen::PopulationI or arrayref of 
Bio::PopGen::IndividualIs
Reference: Weir B.S. (1996) "Genetic Data Analysis II",
Sinauer, Sunderlanm MA.
Methods code
fu_and_li_DdescriptionprevnextTop
sub fu_and_li_D {
     my ($self,$ingroup,$outgroup) = @_;

    my ($seg_sites,$pi,$sample_size,$ancestral,$derived);
    if( ref($ingroup) =~ /ARRAY/i ) {
	$sample_size = scalar @$ingroup;
	# pi - all pairwise differences 
$pi = $self->pi($ingroup); $seg_sites = $self->segregating_sites_count($ingroup); } elsif( ref($ingroup) && $ingroup->isa('Bio::PopGen::PopulationI')) { $sample_size = $ingroup->get_number_individuals; $pi = $self->pi($ingroup); $seg_sites = $self->segregating_sites_count($ingroup); } else { $self->throw("expected an array reference of a list of Bio::PopGen::IndividualI OR a Bio::PopGen::PopulationI object to fu_and_li_D"); return 0; } if( $seg_sites <= 0 ) { $self->warn("mutation total was not > 0, cannot calculate a Fu and Li D"); return 0; } if( ! defined $outgroup ) { $self->warn("Need to provide either an array ref to the outgroup individuals or the number of external mutations"); return 0; } elsif( ref($outgroup) ) { ($ancestral,$derived) = $self->derived_mutations($ingroup,$outgroup); } else { $ancestral = $outgroup; } my $a = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $a += ( 1 / $k );
} my $b = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $b += ( 1 / $k**2 );
} my $c = 2 * ( ( ( $sample_size * $a ) - (2 * ( $sample_size -1 ))) /
( (
$sample_size - 1) * ( $sample_size - 2 ) ) );
my $v = 1 + ( ( $a**2 / ( $b + $a**2 ) ) * ( $c - ( ( $sample_size + 1) / ( $sample_size - 1) ) )); my $u = $a - 1 - $v; my $D = ( $seg_sites - ( $a * $ancestral) ) /
( sqrt ( (
$u * $seg_sites ) + ( $v * $seg_sites **2) ) );
return $D;
}
fu_and_li_D_stardescriptionprevnextTop
sub fu_and_li_D_star {
    my ($self,$individuals) = @_;

    my ($seg_sites,$pi,$sample_size,$singletons);
    if( ref($individuals) =~ /ARRAY/i ) {
	$sample_size = scalar @$individuals;
	# pi - all pairwise differences 
$pi = $self->pi($individuals); $seg_sites = $self->segregating_sites_count($individuals); $singletons = $self->singleton_count($individuals); } elsif( ref($individuals) && $individuals->isa('Bio::PopGen::PopulationI')) { my $pop = $individuals; $sample_size = $pop->get_number_individuals; $pi = $self->pi($pop); $seg_sites = $self->segregating_sites_count($pop); $singletons = $self->singleton_count($pop); } else { $self->throw("expected an array reference of a list of Bio::PopGen::IndividualI OR a Bio::PopGen::PopulationI object to tajima_D"); return 0; } my $a = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $a += ( 1 / $k );
} my $a1 = 0; for(my $k= 1; $k <= $sample_size; $k++ ) { $a1 += ( 1 / $k );
} my $b = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $b += ( 1 / $k**2 );
} my $c = 2 * ( ( ( $sample_size * $a ) - (2 * ( $sample_size -1 ))) /
( (
$sample_size - 1) * ( $sample_size - 2 ) ) );
my $d = $c + ( ($sample_size -2) / ($sample_size - 1)**2 ) +
( 2 /
($sample_size -1) * ( (3/2) - ( (2*$a1 - 3) / ($sample_size -2) ) - ( 1/ $sample_size) )
);
my $v_star = ( ( ($sample_size/($sample_size-1) )**2)*$b + (($a**2)*$d) -
(2*( (
$sample_size*$a*($a+1)) )/(($sample_size-1)**2)) ) /
((
$a**2) + $b);
my $u_star = ( ($sample_size/($sample_size-1))*
(
$a - ($sample_size/ ($sample_size-1)))) - $v_star; my $D_star = ( (($sample_size/($sample_size-1))*$seg_sites) -
(
$a*$singletons) ) / ( sqrt( ($u_star*$seg_sites) + ($v_star*($seg_sites**2)) )); return $D_star;
}
fu_and_li_FdescriptionprevnextTop
sub fu_and_li_F {
    my ($self,$ingroup,$outgroup) = @_;
    my ($seg_sites,$pi,$sample_size,$external,$internal);
    if( ref($ingroup) =~ /ARRAY/i ) {
	$sample_size = scalar @$ingroup;
	# pi - all pairwise differences 
$pi = $self->pi($ingroup); $seg_sites = $self->segregating_sites_count($ingroup); } elsif( ref($ingroup) && $ingroup->isa('Bio::PopGen::PopulationI')) { $sample_size = $ingroup->get_number_individuals; $pi = $self->pi($ingroup); $seg_sites = $self->segregating_sites_count($ingroup); } else { $self->throw("expected an array reference of a list of Bio::PopGen::IndividualI OR a Bio::PopGen::PopulationI object to tajima_D"); return 0; } if( ! defined $outgroup ) { $self->warn("Need to provide either an array ref to the outgroup individuals or the number of external mutations"); return 0; } elsif( ref($outgroup) ) { ($external,$internal) = $self->derived_mutations($ingroup,$outgroup); } else { $external = $outgroup; } my $a = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $a += ( 1 / $k );
} my $a1 = 0; for(my $k= 1; $k <= $sample_size; $k++ ) { $a1 += ( 1 / $k );
} my $b = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $b += ( 1 / $k**2 );
} my $c = 2 * ( ( ( $sample_size * $a ) - (2 * ( $sample_size -1 ))) /
( (
$sample_size - 1) * ( $sample_size - 2 ) ) );
my $v_F = ( $c + ( (2*(($sample_size**2)+$sample_size+3)) /
( (9*
$sample_size)*($sample_size-1) ) ) -
(2/
($sample_size-1)) ) / ( ($a**2)+$b );
my $u_F = ( 1 + ( ($sample_size+1)/(3*($sample_size-1)) )-
( 4*( (
$sample_size+1)/(($sample_size-1)**2) ))* ($a1 - ((2*$sample_size)/($sample_size+1))) ) / ($a - $v_F); my $F = ($pi - $external) / ( sqrt( ($u_F*$seg_sites) +
(
$v_F*($seg_sites**2)) ) );
return $F;
}
fu_and_li_F_stardescriptionprevnextTop
sub fu_and_li_F_star {
    my ($self,$individuals) = @_;

    my ($seg_sites,$pi,$sample_size,$singletons);
    if( ref($individuals) =~ /ARRAY/i ) {
	$sample_size = scalar @$individuals;
	# pi - all pairwise differences 
$pi = $self->pi($individuals); $seg_sites = $self->segregating_sites_count($individuals); $singletons = $self->singleton_count($individuals); } elsif( ref($individuals) && $individuals->isa('Bio::PopGen::PopulationI')) { my $pop = $individuals; $sample_size = $pop->get_number_individuals; $pi = $self->pi($pop); $seg_sites = $self->segregating_sites_count($pop); $singletons = $self->singleton_count($pop); } else { $self->throw("expected an array reference of a list of Bio::PopGen::IndividualI OR a Bio::PopGen::PopulationI object to tajima_D"); return 0; } my $a = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $a += ( 1 / $k );
} my $a1 = 0; for(my $k= 1; $k <= $sample_size; $k++ ) { $a1 += ( 1 / $k );
} my $b = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $b += ( 1 / $k**2 );
} # eq (14)
my $c = 2 * ( (($sample_size * $a) - (2 * ( $sample_size -1 ))) /
((
$sample_size - 1) * ($sample_size - 2)) );
# eq (46)
my $d = $c + ( ($sample_size -2)/ (($sample_size - 1)**2)) +
((2/
($sample_size -1))* ((3/2) - ((2*$a1 - 3)/($sample_size -2)) - (1/$sample_size)));
my $v_F_star = ( $d + ( 2*($sample_size**2+$sample_size+3) /
(9*
$sample_size*($sample_size-1))) -
( (2/
($sample_size-1))* (4*$b - 6 + (8/$sample_size))) )/ ($a**2 + $b); my $u_F_star = ( ($sample_size / ($sample_size-1)) +
((
$sample_size+1)/(3*($sample_size-1))) - ( 2 * (2 / ($sample_size * ($sample_size-1)))) +
(2*( (
$sample_size+1)/($sample_size-1)**2)* ($a1 - ((2*$sample_size)/($sample_size+1))) )) / ($a - $v_F_star); my $F_star = ( $pi - (( ($sample_size-1)/ $sample_size)*$singletons)) / sqrt ( ($u_F_star*$seg_sites) + ($v_F_star*($seg_sites**2))); return $F_star;
}
tajima_DdescriptionprevnextTop
sub tajima_D {
    my ($self,$individuals) = @_;
    my ($seg_sites,$pi,$sample_size);

    if( ref($individuals) =~ /ARRAY/i ) {
	$sample_size = scalar @$individuals;
	# pi - all pairwise differences 
$pi = $self->pi($individuals); $seg_sites = $self->segregating_sites_count($individuals); } elsif( ref($individuals) && $individuals->isa('Bio::PopGen::PopulationI')) { my $pop = $individuals; $sample_size = $pop->get_number_individuals; $pi = $self->pi($pop); $seg_sites = $self->segregating_sites_count($pop); } else { $self->throw("expected an array reference of a list of Bio::PopGen::IndividualI OR a Bio::PopGen::PopulationI object to tajima_D"); return 0; } my $a1 = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $a1 += ( 1 / $k );
} my $a2 = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $a2 += ( 1 / $k**2 );
} my $b1 = ( $sample_size + 1 ) / ( 3* ( $sample_size - 1) );
my $b2 = ( 2 * ( $sample_size ** 2 + $sample_size + 3) ) /
( ( 9 *
$sample_size) * ( $sample_size - 1) );
my $c1 = $b1 - ( 1 / $a1 );
my $c2 = $b2 - ( ( $sample_size + 2 ) /
(
$a1 * $sample_size))+( $a2 / $a1 ** 2); my $e1 = $c1 / $a1;
my $e2 = $c2 / ( $a1**2 + $a2 );
my $D = ( $pi - ( $seg_sites / $a1 ) ) / sqrt ( ($e1 * $seg_sites) + (( $e2 * $seg_sites) * ( $seg_sites - 1))); return $D;
}
pidescriptionprevnextTop
sub pi {
    my ($self,$individuals,$numsites) = @_;
    my (%data,@marker_names,$sample_size);

    if( ref($individuals) =~ /ARRAY/i ) {
	# one possible argument is an arrayref of Bio::PopGen::IndividualI objs
@marker_names = $individuals->[0]->get_marker_names; $sample_size = scalar @$individuals; # Here we're calculating the allele frequencies
my %marker_total; foreach my $ind ( @$individuals ) { if( ! $ind->isa('Bio::PopGen::IndividualI') ) { $self->warn("Expected an arrayref of Bio::PopGen::IndividualI objects, this is a ".ref($ind)."\n"); return 0; } foreach my $m ( @marker_names ) { foreach my $allele (map { $_->get_Alleles} $ind->get_Genotypes($m) ) { $data{$m}->{$allele}++; $marker_total{$m}++; } } } while( my ($marker,$count) = each %marker_total ) { foreach my $c ( values %{$data{$marker}} ) { $c /= $count;
} } # %data will contain allele frequencies for each marker, allele
} elsif( ref($individuals) && $individuals->isa('Bio::PopGen::PopulationI') ) { my $pop = $individuals; $sample_size = $pop->get_number_individuals; foreach my $marker( $pop->get_Markers ) { push @marker_names, $marker->name; $data{$marker->name} = {$marker->get_Allele_Frequencies}; } } else { $self->throw("expected an array reference of a list of Bio::PopGen::IndividualI to pi"); } # doing all pairwise combinations
# For now we assume that all individuals have the same markers
my ($diffcount,$totalcompare) = (0,0); my $pi = 0; foreach my $markerdat ( values %data ) { my $totalalleles; # this will only be different among markers
# when there is missing data
my @alleles = keys %$markerdat; foreach my $al ( @alleles ) { $totalalleles += $markerdat->{$al} } for( my $i =0; $i < scalar @alleles -1; $i++ ) { my ($a1,$a2) = ( $alleles[$i], $alleles[$i+1]); $pi += $self->heterozygosity($sample_size, $markerdat->{$a1} / $totalalleles,
$markerdat->{$a2} / $totalalleles); } } $self->debug( "pi=$pi\n"); if( $numsites ) { return $pi / $numsites;
} else { return $pi; }
}
thetadescriptionprevnextTop
sub theta {
    my $self = shift;    
    my ( $sample_size, $seg_sites,$totalsites) = @_;
    if( ref($sample_size) =~ /ARRAY/i ) {
	my $samps = $sample_size;
	$totalsites = $seg_sites; # only 2 arguments if one is an array
my %data; my @marker_names = $samps->[0]->get_marker_names; # we need to calculate number of polymorphic sites
$seg_sites = $self->segregating_sites_count($samps); $sample_size = scalar @$samps; } elsif(ref($sample_size) && $sample_size->isa('Bio::PopGen::PopulationI') ) { # This will handle the case when we pass in a PopulationI object
my $pop = $sample_size; $totalsites = $seg_sites; # shift the arguments over by one
$sample_size = $pop->get_number_individuals; $seg_sites = $self->segregating_sites_count($pop); } my $a1 = 0; for(my $k= 1; $k < $sample_size; $k++ ) { $a1 += ( 1 / $k );
} if( $totalsites ) { # 0 and undef are the same can't divide by them
$seg_sites /= $totalsites;
} return $seg_sites / $a1;
}
singleton_countdescriptionprevnextTop
sub singleton_count {
    my ($self,$individuals) = @_;

    my @inds;
    if( ref($individuals) =~ /ARRAY/ ) {
	@inds = @$individuals;
    } elsif( ref($individuals) && 
	     $individuals->isa('Bio::PopGen::PopulationI') ) {
	my $pop = $individuals;
	@inds = $pop->get_Individuals();
	unless( @inds ) { 
	    $self->warn("Need to provide a population which has individuals loaded, not just a population with allele frequencies");
	    return 0;
	}
    } else {
	$self->warn("Expected either a PopulationI object or an arrayref of IndividualI objects");
	return 0;
    }
    # find number of sites where a particular allele is only seen once
my ($singleton_allele_ct,%sites) = (0); # first collect all the alleles into a hash structure
foreach my $n ( @inds ) { if( ! $n->isa('Bio::PopGen::IndividualI') ) { $self->warn("Expected an arrayref of Bio::PopGen::IndividualI objects, this is a ".ref($n)."\n"); return 0; } foreach my $g ( $n->get_Genotypes ) { my ($nm,@alleles) = ($g->marker_name, $g->get_Alleles); foreach my $allele (@alleles ) { $sites{$nm}->{$allele}++; } } } foreach my $site ( values %sites ) { # don't really care what the name is
foreach my $allelect ( values %$site ) { #
# find the sites which have an allele with only 1 copy
$singleton_allele_ct++ if( $allelect == 1 ); } } return $singleton_allele_ct;
}
segregating_sites_countdescriptionprevnextTop
sub segregating_sites_count {
   my ($self,$individuals) = @_;
   my $type = ref($individuals);
   my $seg_sites = 0;
   if( $type =~ /ARRAY/i ) {
       my %sites;
       foreach my $n ( @$individuals ) {
	   if( ! $n->isa('Bio::PopGen::IndividualI') ) {
	       $self->warn("Expected an arrayref of Bio::PopGen::IndividualI objects, this is a ".ref($n)."\n");
	       return 0;
	   }
	   foreach my $g ( $n->get_Genotypes ) {
	       my ($nm,@alleles) = ($g->marker_name, $g->get_Alleles);
	       foreach my $allele (@alleles ) {
		   $sites{$nm}->{$allele}++;
	       }
	   }
       }
       foreach my $site ( values %sites ) { # use values b/c we don't 
# really care what the name is
# find the sites which >1 allele
$seg_sites++ if( keys %$site > 1 ); } } elsif( $type && $individuals->isa('Bio::PopGen::PopulationI') ) { foreach my $marker ( $individuals->get_Markers ) { my @alleles = $marker->get_Alleles; $seg_sites++ if ( scalar @alleles > 1 ); } } else { $self->warn("segregating_sites_count expects either a PopulationI object or a list of IndividualI objects"); return 0; } return $seg_sites;
}
heterozygositydescriptionprevnextTop
sub heterozygosity {
    my ($self,$samp_size, $freq1,$freq2) = @_;
    if( ! $freq2 ) { $freq2 = 1 - $freq1 }
    if( $freq1 > 1 || $freq2 > 1 ) { 
	$self->warn("heterozygosity expects frequencies to be less than 1");
    }
    my $sum = ($freq1**2) + (($freq2)**2);
    my $h = ( $samp_size*(1- $sum) ) / ($samp_size - 1) ;
return $h;
}
derived_mutationsdescriptionprevnextTop
sub derived_mutations {
   my ($self,$ingroup,$outgroup) = @_;
   my (%indata,%outdata,@marker_names);

   # basically we have to do some type checking
# if that perl were typed...
my ($itype,$otype) = (ref($ingroup),ref($outgroup)); return $outgroup unless( $otype ); # we expect arrayrefs or objects, nums
# are already the value we
# are searching for
# pick apart the ingroup
# get the data
if( ref($ingroup) =~ /ARRAY/i ) { if( ! ref($ingroup->[0]) || ! $ingroup->[0]->isa('Bio::PopGen::IndividualI') ) { $self->warn("Expected an arrayref of Bio::PopGen::IndividualI objects or a Population for ingroup in external_mutations"); return 0; } # we assume that all individuals have the same markers
# i.e. that they are aligned
@marker_names = $ingroup->[0]->get_marker_names; for my $ind ( @$ingroup ) { for my $m ( @marker_names ) { for my $allele ( map { $_->get_Alleles } $ind->get_Genotypes($m) ) { $indata{$m}->{$allele}++; } } } } elsif( ref($ingroup) && $ingroup->isa('Bio::PopGen::PopulationI') ) { @marker_names = $ingroup->get_marker_names; for my $ind ( $ingroup->get_Individuals() ) { for my $m ( @marker_names ) { for my $allele ( map { $_->get_Alleles} $ind->get_Genotypes($m) ) { $indata{$m}->{$allele}++; } } } } else { $self->warn("Need an arrayref of Bio::PopGen::IndividualI objs or a Bio::PopGen::Population for ingroup in external_mutations"); return 0; } if( $otype =~ /ARRAY/i ) { if( ! ref($outgroup->[0]) || ! $outgroup->[0]->isa('Bio::PopGen::IndividualI') ) { $self->warn("Expected an arrayref of Bio::PopGen::IndividualI objects or a Population for outgroup in external_mutations"); return 0; } for my $ind ( @$outgroup ) { for my $m ( @marker_names ) { for my $allele ( map { $_->get_Alleles } $ind->get_Genotypes($m) ) { $outdata{$m}->{$allele}++; } } } } elsif( $otype->isa('Bio::PopGen::PopulationI') ) { for my $ind ( $outgroup->get_Individuals() ) { for my $m ( @marker_names ) { for my $allele ( map { $_->get_Alleles} $ind->get_Genotypes($m) ) { $outdata{$m}->{$allele}++; } } } } elsif( $otype->isa('Bio::PopGen::PopulationI') ) { $self->warn("Need an arrayref of Bio::PopGen::IndividualI objs or a Bio::PopGen::Population for outgroup in external_mutations"); return 0; } # derived mutations are defined as
#
# ingroup (G A T)
# outgroup (A)
# derived mutations are G and T, A is the external mutation
# ingroup (A T)
# outgroup (C)
# derived mutations A,T no external/ancestral mutations
# ingroup (G A T)
# outgroup (A T)
# cannot determine
my ($internal,$external); foreach my $marker ( @marker_names ) { my @outalleles = keys %{$outdata{$marker}}; my @in_alleles = keys %{$indata{$marker}}; next if( @outalleles > 1 || @in_alleles == 1); for my $allele ( @in_alleles ) { if( ! exists $outdata{$marker}->{$allele} ) { if( $indata{$marker}->{$allele} == 1 ) { $external++; } else { $internal++; } } } } return ($external, $internal);
}
composite_LDdescriptionprevnextTop
sub composite_LD {
    my ($self,$pop) = @_;
    if( ref($pop) =~ /ARRAY/i ) {
	if( ref($pop->[0]) && $pop->[0]->isa('Bio::PopGen::IndividualI') ) {
	    $pop = new Bio::PopGen::Population(-individuals => @$pop);
	} else { 
	    $self->warn("composite_LD expects a Bio::PopGen::PopulationI or an arrayref of Bio::PopGen::IndividualI objects");
	    return ();
	}
    } elsif( ! ref($pop) || ! $pop->isa('Bio::PopGen::PopulationI') ) {
	$self->warn("composite_LD expects a Bio::PopGen::PopulationI or an arrayref of Bio::PopGen::IndividualI objects");
	return ();
    }

    my @marker_names = $pop->get_marker_names;
    my @inds = $pop->get_Individuals;
    my $num_inds = scalar @inds;
    my (%lookup);
    # calculate allele frequencies for each marker from the population
# use the built-in get_Marker to get the allele freqs
# we still need to calculate the genotype frequencies
foreach my $marker_name ( @marker_names ) { my(%allelef); foreach my $ind ( @inds ) { my ($genotype) = $ind->get_Genotypes(-marker => $marker_name); my @alleles = sort $genotype->get_Alleles; next if( scalar @alleles != 2); my $genostr = join(',', @alleles); $allelef{$alleles[0]}++; $allelef{$alleles[1]}++; } # we should check for cases where there > 2 alleles or
# only 1 allele and throw out those markers.
my @alleles = sort keys %allelef; my $allele_count = scalar @alleles; # test if site is polymorphic
if( $allele_count != 2) { # only really warn if we're seeing multi-allele
$self->warn("Skipping $marker_name because it has $allele_count alleles (".join(',',@alleles)."),\n composite_LD will currently only work for biallelic markers") if $allele_count > 2; next; # skip this marker
} # Need to do something here to detect alleles which aren't
# a single character
if( length($alleles[0]) != 1 || length($alleles[1]) != 1 ) { $self->warn("An individual has an allele which is not a single base, this is currently not supported in composite_LD - consider recoding the allele as a single character"); next; } # fix the call for allele 1 (A or B) and
# allele 2 (a or b) in terms of how we'll do the
# N square from Weir p.126
$self->debug( "$alleles[0] is 1, $alleles[1] is 2 for $marker_name\n"); $lookup{$marker_name}->{'1'} = $alleles[0]; $lookup{$marker_name}->{'2'} = $alleles[1]; } @marker_names = sort keys %lookup; my $site_count = scalar @marker_names; # where the final data will be stored
my %stats_for_sites; # standard way of generating pairwise combos
# LD is done by comparing all the pairwise site (marker)
# combinations and keeping track of the genotype and
# pairwise genotype (ie genotypes of the 2 sites) frequencies
for( my $i = 0; $i < $site_count - 1; $i++ ) { my $site1 = $marker_names[$i]; my (%genotypes, %total_genotype_count, %total_pairwisegeno_count,%pairwise_genotypes); for( my $j = $i+1; $j < $site_count ; $j++) { my (%genotypes, %total_genotype_count, %total_pairwisegeno_count,%pairwise_genotypes); my $site2 = $marker_names[$j]; my (%allele_count,%allele_freqs) = (0,0); foreach my $ind ( @inds ) { # build string of genotype at site 1
my ($genotype1) = $ind->get_Genotypes(-marker => $site1); my @alleles1 = sort $genotype1->get_Alleles; # if an individual has only one available allele
# (has a blank or N for one of the chromosomes)
# we don't want to use it in our calculation
next unless( scalar @alleles1 == 2); my $genostr1 = join(',', @alleles1); # build string of genotype at site 2
my ($genotype2) = $ind->get_Genotypes(-marker => $site2); my @alleles2 = sort $genotype2->get_Alleles; my $genostr2 = join(',', @alleles2); next unless( scalar @alleles2 == 2); for (@alleles1) { $allele_count{$site1}++; $allele_freqs{$site1}->{$_}++; } $genotypes{$site1}->{$genostr1}++; $total_genotype_count{$site1}++; for (@alleles2) { $allele_count{$site2}++; $allele_freqs{$site2}->{$_}++; } $genotypes{$site2}->{$genostr2}++; $total_genotype_count{$site2}++; # We are using the $site1,$site2 to signify
# a unique key
$pairwise_genotypes{"$site1,$site2"}->{"$genostr1,$genostr2"}++; # some individuals
$total_pairwisegeno_count{"$site1,$site2"}++; } for my $site ( %allele_freqs ) { for my $al ( keys %{ $allele_freqs{$site} } ) { $allele_freqs{$site}->{$al} /= $allele_count{$site};
} } my $n = $total_pairwisegeno_count{"$site1,$site2"}; # number of inds
# 'A' and 'B' are two loci or in our case site1 and site2
my $allele1_site1 = $lookup{$site1}->{'1'}; # this is the BigA allele
my $allele1_site2 = $lookup{$site2}->{'1'}; # this is the BigB allele
my $allele2_site1 = $lookup{$site1}->{'2'}; # this is the LittleA allele
my $allele2_site2 = $lookup{$site2}->{'2'}; # this is the LittleB allele
# AABB
my $N1genostr = join(",",( $allele1_site1, $allele1_site1, $allele1_site2, $allele1_site2)); $self->debug(" [$site1,$site2](AABB) N1genostr=$N1genostr\n"); # AABb
my $N2genostr = join(",",( $allele1_site1, $allele1_site1, $allele1_site2, $allele2_site2)); $self->debug(" [$site1,$site2](AABb) N2genostr=$N2genostr\n"); # AaBB
my $N4genostr = join(",",( $allele1_site1, $allele2_site1, $allele1_site2, $allele1_site2)); $self->debug(" [$site1,$site2](AaBB) N4genostr=$N4genostr\n"); # AaBb
my $N5genostr = join(",",( $allele1_site1, $allele2_site1, $allele1_site2, $allele2_site2)); $self->debug(" [$site1,$site2](AaBb) N5genostr=$N5genostr\n"); # count of AABB in
my $n1 = $pairwise_genotypes{"$site1,$site2"}->{$N1genostr} || 0; # count of AABb in
my $n2 = $pairwise_genotypes{"$site1,$site2"}->{$N2genostr} || 0; # count of AaBB in
my $n4 = $pairwise_genotypes{"$site1,$site2"}->{$N4genostr} || 0; # count of AaBb in
my $n5 = $pairwise_genotypes{"$site1,$site2"}->{$N5genostr} || 0; my $homozA_site1 = join(",", ($allele1_site1,$allele1_site1)); my $homozB_site2 = join(",", ($allele1_site2,$allele1_site2)); my $p_AA = ($genotypes{$site1}->{$homozA_site1} || 0) / $n;
my $p_BB = ($genotypes{$site2}->{$homozB_site2} || 0) / $n;
my $p_A = $allele_freqs{$site1}->{$allele1_site1} || 0; # an individual allele freq
my $p_a = 1 - $p_A; my $p_B = $allele_freqs{$site2}->{$allele1_site2} || 0; # an individual allele freq
my $p_b = 1 - $p_B; # variance of allele frequencies
my $pi_A = $p_A * $p_a; my $pi_B = $p_B * $p_b; # hardy weinberg
my $D_A = $p_AA - $p_A**2; my $D_B = $p_BB - $p_B**2; my $n_AB = 2*$n1 + $n2 + $n4 + 0.5 * $n5; $self->debug("n_AB=$n_AB -- n1=$n1, n2=$n2 n4=$n4 n5=$n5\n"); my $delta_AB = (1 / $n ) * ( $n_AB ) - ( 2 * $p_A * $p_B );
$self->debug("delta_AB=$delta_AB -- n=$n, n_AB=$n_AB p_A=$p_A, p_B=$p_B\n"); $self->debug(sprintf(" (%d * %.4f) / ( %.2f + %.2f) * ( %.2f + %.2f)\n ", $n,$delta_AB**2, $pi_A, $D_A, $pi_B, $D_B)); my $chisquared; eval { $chisquared = ( $n * ($delta_AB**2) ) /
( (
$pi_A + $D_A) * ( $pi_B + $D_B) );
}; if( $@ ) { $self->debug("Skipping the site because the denom is 0.\nsite1=$site1, site2=$site2 : pi_A=$pi_A, pi_B=$pi_B D_A=$D_A, D_B=$D_B\n"); next; } # this will be an upper triangular matrix
$stats_for_sites{$site1}->{$site2} = [$delta_AB,$chisquared]; } } return %stats_for_sites;
}
General documentation
FEEDBACKTop
Mailing ListsTop
User feedback is an integral part of the evolution of this and other
Bioperl modules. Send your comments and suggestions preferably to
the Bioperl mailing list. Your participation is much appreciated.
  bioperl-l@bioperl.org              - General discussion
  http://bioperl.org/MailList.shtml  - About the mailing lists
Reporting BugsTop
Report bugs to the Bioperl bug tracking system to help us keep track
of the bugs and their resolution. Bug reports can be submitted via
the web:
  http://bugzilla.bioperl.org/
AUTHOR - Jason Stajich, Matthew HahnTop
Email jason-at-bioperl-dot-org
Matt Hahn <matthew.hahn-at-duke.duke>
CONTRIBUTORSTop
Additional contributors names and emails here
APPENDIXTop
The rest of the documentation details each of the object methods.
Internal methods are usually preceded with a _
newTop
 Title   : new
 Usage   : my $obj = new Bio::PopGen::Statistics();
 Function: Builds a new Bio::PopGen::Statistics object 
 Returns : an instance of Bio::PopGen::Statistics
 Args    : none