Bio::EnsEMBL::Compara::RunnableDB NJTREE_PHYML
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Summary
Bio::EnsEMBL::Compara::RunnableDB::NJTREE_PHYML
Package variables
No package variables defined.
Included modules
Bio::AlignIO
Bio::EnsEMBL::Compara::DBSQL::DBAdaptor
Bio::EnsEMBL::Compara::Graph::NewickParser
Bio::EnsEMBL::Compara::Member
Bio::EnsEMBL::DBSQL::DBAdaptor
Bio::EnsEMBL::Hive
Bio::SimpleAlign
File::Basename
Getopt::Long
IO::File
Time::HiRes qw ( time gettimeofday tv_interval )
Inherit
Bio::EnsEMBL::Hive::Process
Synopsis
my $db = Bio::EnsEMBL::Compara::DBAdaptor->new($locator);
my $njtree_phyml = Bio::EnsEMBL::Compara::RunnableDB::NJTREE_PHYML->new
(
-db => $db,
-input_id => $input_id,
-analysis => $analysis
);
$njtree_phyml->fetch_input(); #reads from DB
$njtree_phyml->run();
$njtree_phyml->output();
$njtree_phyml->write_output(); #writes to DB
Description
This Analysis/RunnableDB is designed to take ProteinTree as input
This must already have a multiple alignment run on it. It uses that alignment
as input into the NJTREE PHYML program which then generates a phylogenetic tree
input_id/parameters format eg: "{'protein_tree_id'=>1234}"
protein_tree_id : use 'id' to fetch a cluster from the ProteinTree
Methods
DESTROY
No description
Code
_store_tree_tags
No description
Code
check_job_fail_options
No description
Code
dumpTreeMultipleAlignmentToWorkdir
No description
Code
fetch_inputDescriptionCode
get_params
No description
Code
parse_newick_into_proteintree
No description
Code
print_params
No description
Code
runDescriptionCode
run_njtree_phyml
No description
Code
store_proteintree
No description
Code
store_tags
No description
Code
write_outputDescriptionCode
Methods description
fetch_inputcode    nextTop
    Title   :   fetch_input
Usage : $self->fetch_input
Function: Fetches input data for repeatmasker from the database
Returns : none
Args : none
runcodeprevnextTop
    Title   :   run
Usage : $self->run
Function: runs NJTREE PHYML
Returns : none
Args : none
write_outputcodeprevnextTop
    Title   :   write_output
Usage : $self->write_output
Function: stores proteintree
Returns : none
Args : none
Methods code
DESTROYdescriptionprevnextTop
sub DESTROY {
  my $self = shift;

  if($self->{'protein_tree'}) {
    printf("NJTREE_PHYML::DESTROY  releasing tree\n") if($self->debug);
    $self->{'protein_tree'}->release_tree;
    $self->{'protein_tree'} = undef;
  }

  $self->SUPER::DESTROY if $self->can("SUPER::DESTROY");
}


##########################################
#
# internal methods
#
##########################################
}
_store_tree_tagsdescriptionprevnextTop
sub _store_tree_tags {
    my $self = shift;
    my $tree = $self->{'protein_tree'};
    my $pta = $self->{'comparaDBA'}->get_ProteinTreeAdaptor;

    print "Storing Tree tags...\n";
    $tree->_load_tags();

    my @leaves = @{$tree->get_all_leaves};
    my @nodes = @{$tree->get_all_nodes};

    # Node is a root node (full tree).
my $node_is_root = 0; my @roots = @{$pta->fetch_all_roots()}; foreach my $root (@roots) { $node_is_root = 1 if ($root->node_id == $tree->parent->node_id); } $tree->store_tag("node_is_root",$node_is_root); # Node is leaf node.
my $node_is_leaf = 0; $node_is_leaf = 1 if ($tree->is_leaf); $tree->store_tag("node_is_leaf",$node_is_leaf); # Tree number of leaves.
my $tree_num_leaves = scalar(@leaves); $tree->store_tag("tree_num_leaves",$tree_num_leaves); # Tree number of human peptides contained.
my $num_hum_peps = 0; foreach my $leaf (@leaves) { $num_hum_peps++ if ($leaf->taxon_id == 9606); } $tree->store_tag("tree_num_human_peps",$num_hum_peps); # Tree max root-to-tip distance.
my $tree_max_length = $tree->max_distance; $tree->store_tag("tree_max_length",$tree_max_length); # Tree max single branch length.
my $tree_max_branch = 0; foreach my $node (@nodes) { my $dist = $node->distance_to_parent; $tree_max_branch = $dist if ($dist > $tree_max_branch); } $tree->store_tag("tree_max_branch",$tree_max_branch); # Tree number of duplications and speciations.
my $tree_num_leaves = scalar(@{$tree->get_all_leaves}); my $num_dups = 0; my $num_specs = 0; foreach my $node (@{$tree->get_all_nodes}) { my $dup = $node->get_tagvalue("Duplication"); if ($dup ne '' && $dup > 0) { $num_dups++; } else { $num_specs++; } } $tree->store_tag("tree_num_dup_nodes",$num_dups); $tree->store_tag("tree_num_spec_nodes",$num_specs); print "Done storing stuff!\n" if ($self->debug); } 1;
}
check_job_fail_optionsdescriptionprevnextTop
sub check_job_fail_options {
  my $self = shift;

  if ($self->input_job->retry_count == 1) {
    if ($self->{'protein_tree'}->get_tagvalue('gene_count') > 200 && !defined($self->worker->{HIGHMEM})) {
      $self->input_job->adaptor->reset_highmem_job_by_dbID($self->input_job->dbID);
      $self->DESTROY;
      throw("NJTREE PHYML job too big: try something else and FAIL it");
    }
  }

  # This will go to QuickTreeBreak
if($self->input_job->retry_count >= 2 && !defined($self->{'jackknife'})) { $self->dataflow_output_id($self->input_id, 3); $self->input_job->update_status('FAILED'); $self->DESTROY; throw("NJTREE PHYML job failed >=3 times: try something else and FAIL it"); } } ########################################################
#
# ProteinTree input/output section
#
########################################################
}
dumpTreeMultipleAlignmentToWorkdirdescriptionprevnextTop
sub dumpTreeMultipleAlignmentToWorkdir {
  my $self = shift;
  my $tree = shift;

  my $leafcount = scalar(@{$tree->get_all_leaves});
  if($leafcount<3) {
    printf(STDERR "tree cluster %d has <3 proteins - can not build a tree\n", 
           $tree->node_id);
    return undef;
  }

  $self->{'file_root'} = 
    $self->worker_temp_directory. "proteintree_". $tree->node_id;
  $self->{'file_root'} =~ s/\/\//\//g;  # converts any // in path to /
my $aln_file = $self->{'file_root'} . ".aln"; return $aln_file if(-e $aln_file); if($self->debug) { printf("dumpTreeMultipleAlignmentToWorkdir : %d members\n", $leafcount); print("aln_file = '$aln_file'\n"); } open(OUTSEQ, ">$aln_file") or $self->throw("Error opening $aln_file for write"); # Using append_taxon_id will give nice seqnames_taxonids needed for
# njtree species_tree matching
my %sa_params = ($self->{use_genomedb_id}) ? ('-APPEND_GENOMEDB_ID', 1) : ('-APPEND_TAXON_ID', 1); my $sa = $tree->get_SimpleAlign ( -id_type => 'MEMBER', -cdna=>$self->{'cdna'}, -stop2x => 1, %sa_params ); $sa->set_displayname_flat(1); if (defined($self->{'jackknife'})) { # my $coverage_hash;
my $empty_hash; foreach my $seq ($sa->each_seq) { my $sequence = $seq->seq; $sequence =~ s/\-//g; my $full_length = length($sequence); $sequence =~ s/N//g; my $covered_length = length($sequence); my $coverage_proportion = $covered_length/$full_length;
my $empty_length = $full_length - $covered_length; # $coverage_hash->{$coverage_proportion} = $seq->display_id if ($coverage_proportion < 1);
$empty_hash->{$empty_length} = $seq->display_id if ($empty_length > 0); } my @lowest = sort {$b<=>$a} keys %$empty_hash; my $i = 0; while ($i < $self->{jackknife}) { $sa->remove_seq($sa->each_seq_with_id($empty_hash->{$lowest[$i]})); $sa = $sa->remove_gaps(undef,1); $i++; } } my $alignIO = Bio::AlignIO->newFh ( -fh =>\* OUTSEQ, -format => "fasta" ); print $alignIO $sa; close OUTSEQ; $self->{'input_aln'} = $aln_file; return $aln_file;
}
fetch_inputdescriptionprevnextTop
sub fetch_input {
  my( $self) = @_;

  $self->{'cdna'}           = 1;
  $self->{'bootstrap'}      = 1;
  $self->{'max_gene_count'} = 200;

  $self->throw("No input_id") unless defined($self->input_id);

  #create a Compara::DBAdaptor which shares the same DBI handle
#with the pipeline DBAdaptor that is based into this runnable
$self->{'comparaDBA'} = Bio::EnsEMBL::Compara::DBSQL::DBAdaptor->new ( -DBCONN=>$self->db->dbc ); $self->get_params($self->parameters); $self->get_params($self->input_id); $self->print_params if($self->debug); $self->check_job_fail_options; $self->check_if_exit_cleanly; unless($self->{'protein_tree'}) { throw("undefined ProteinTree as input\n"); } if ($self->{'protein_tree'}->get_tagvalue('gene_count') > $self->{'max_gene_count'}) { # 3 is QuickTreeBreak -- 2 is NJTREE_PHYML jackknife
$self->dataflow_output_id($self->input_id, 3); $self->input_job->update_status('FAILED'); $self->{'protein_tree'}->release_tree; $self->{'protein_tree'} = undef; throw("NJTREE_PHYML : cluster size over threshold and FAIL it"); } return 1;
}
get_paramsdescriptionprevnextTop
sub get_params {
  my $self         = shift;
  my $param_string = shift;

  return unless($param_string);
  print("parsing parameter string : ",$param_string,"\n") if($self->debug);

  my $params = eval($param_string);
  return unless($params);

  if (defined $params->{'njtree_phyml_analysis_data_id'}) {
    my $njtree_phyml_analysis_data_id = $params->{'njtree_phyml_analysis_data_id'};
    my $ada = $self->db->get_AnalysisDataAdaptor;
    my $new_params = eval($ada->fetch_by_dbID($njtree_phyml_analysis_data_id));
    if (defined $new_params) {
      $params = $new_params;
    }
  }

  if($self->debug) {
    foreach my $key (keys %$params) {
      print("  $key : ", $params->{$key}, "\n");
    }
  }

  if(defined($params->{'protein_tree_id'})) {
    $self->{'protein_tree'} =  
         $self->{'comparaDBA'}->get_ProteinTreeAdaptor->
         fetch_node_by_node_id($params->{'protein_tree_id'});
  }
  $self->{'jackknife'} = $params->{'jackknife'} if(defined($params->{'jackknife'}));
  $self->{'cdna'} = $params->{'cdna'} if(defined($params->{'cdna'}));
  $self->{'max_gene_count'} = 
    $params->{'max_gene_count'} if(defined($params->{'max_gene_count'}));

  foreach my $key (qw[cdna max_gene_count species_tree_file honeycomb_dir use_genomedb_id]) {
    my $value = $params->{$key};
    $self->{$key} = $value if defined $value;
  }

  return;
}
parse_newick_into_proteintreedescriptionprevnextTop
sub parse_newick_into_proteintree {
  my $self = shift;
  my $newick_file =  $self->{'newick_file'};
  my $tree = $self->{'protein_tree'};
  
  #cleanup old tree structure- 
# flatten and reduce to only AlignedMember leaves
$tree->flatten_tree; $tree->print_tree(20) if($self->debug); foreach my $node (@{$tree->get_all_leaves}) { next if($node->isa('Bio::EnsEMBL::Compara::AlignedMember')); $node->disavow_parent; } #parse newick into a new tree object structure
my $newick = ''; print("load from file $newick_file\n") if($self->debug); open (FH, $newick_file) or throw("Couldnt open newick file [$newick_file]"); while(<FH>) { $newick .= $_; } close(FH); my $newtree = Bio::EnsEMBL::Compara::Graph::NewickParser::parse_newick_into_tree($newick); $newtree->print_tree(20) if($self->debug > 1); # get rid of the taxon_id needed by njtree -- name tag
foreach my $leaf (@{$newtree->get_all_leaves}) { my $njtree_phyml_name = $leaf->get_tagvalue('name'); $njtree_phyml_name =~ /(\d+)\_\d+/; my $member_name = $1; $leaf->add_tag('name', $member_name); } # Leaves of newick tree are named with member_id of members from
# input tree move members (leaves) of input tree into newick tree to
# mirror the 'member_id' nodes
foreach my $member (@{$tree->get_all_leaves}) { my $tmpnode = $newtree->find_node_by_name($member->member_id); if($tmpnode) { $tmpnode->add_child($member, 0.0); $tmpnode->minimize_node; #tmpnode is now redundant so it is removed
} else { print("unable to find node in newick for member"); $member->print_member; } } # Merge the trees so that the children of the newick tree are now
# attached to the input tree's root node
$tree->merge_children($newtree); # Newick tree is now empty so release it
$newtree->release_tree; $tree->print_tree if($self->debug); # check here on the leaf to test if they all are AlignedMembers as
# minimize_tree/minimize_node might not work properly
foreach my $leaf (@{$self->{'protein_tree'}->get_all_leaves}) { unless($leaf->isa('Bio::EnsEMBL::Compara::AlignedMember')) { throw("Phyml tree does not have all leaves as AlignedMember\n"); } } return undef;
}
print_paramsdescriptionprevnextTop
sub print_params {
  my $self = shift;

  print("params:\n");
  print("  tree_id   : ", $self->{'protein_tree'}->node_id,"\n") if($self->{'protein_tree'});
  print("  cdna      : ", $self->{'cdna'},"\n");
}
rundescriptionprevnextTop
sub run {
  my $self = shift;
  $self->check_if_exit_cleanly;
  $self->run_njtree_phyml;
}
run_njtree_phymldescriptionprevnextTop
sub run_njtree_phyml {
  my $self = shift;

  my $starttime = time()*1000;

  $self->{'cdna'} = 1; #always use cdna for njtree_phyml
$self->{'input_aln'} = $self->dumpTreeMultipleAlignmentToWorkdir ( $self->{'protein_tree'} ); return unless($self->{'input_aln'}); $self->{'newick_file'} = $self->{'input_aln'} . "_njtree_phyml_tree.txt "; my $njtree_phyml_executable = $self->analysis->program_file || ''; unless (-e $njtree_phyml_executable) { if (`uname -m` =~ /ia64/) { $njtree_phyml_executable = "/nfs/acari/avilella/src/treesoft/trunk/treebest/ia64/treebest"; } else { # it is a linux machine
# md5sum 91a9da7ad7d38ebedd5ce363a28d509b
# $njtree_phyml_executable = "/lustre/work1/ensembl/avilella/bin/i386/njtree_gcc";
# $njtree_phyml_executable = "/nfs/acari/avilella/src/_treesoft/treebest/treebest";
$njtree_phyml_executable = "/nfs/acari/avilella/src/treesoft/trunk/treebest/treebest"; } } throw("can't find a njtree executable to run\n") unless(-e $njtree_phyml_executable); throw("can't find species_tree_file\n") unless(-e $self->{'species_tree_file'}); # ./njtree best -f spec-v4.1.nh -p tree -o $BASENAME.best.nhx \
# $BASENAME.nucl.mfa -b 100 2>&1/dev/null
my $cmd = $njtree_phyml_executable; if (1 == $self->{'bootstrap'}) { $cmd .= " best "; if (defined($self->{'species_tree_file'})) { $cmd .= " -f ". $self->{'species_tree_file'}; } $cmd .= " ". $self->{'input_aln'}; $cmd .= " -p tree "; $cmd .= " -o " . $self->{'newick_file'}; my $logfile = $self->worker_temp_directory. "proteintree_". $self->{'protein_tree'}->node_id . ".log"; my $errfile = $self->worker_temp_directory. "proteintree_". $self->{'protein_tree'}->node_id . ".err"; $cmd .= " 1>$logfile 2>$errfile"; # $cmd .= " 2>&1 > /dev/null" unless($self->debug);
$self->{'comparaDBA'}->dbc->disconnect_when_inactive(1); print("$cmd\n") if($self->debug); my $worker_temp_directory = $self->worker_temp_directory; unless(system("cd $worker_temp_directory; $cmd") == 0) { print("$cmd\n"); open ERRFILE,"$errfile" or die "couldnt open logfile $errfile: $!\n"; while (<ERRFILE>) { if ($_ =~ /NNI/) { # Do jack-knife treebest starting by the sequence with more Ns
my $jackknife_value = $self->{'jackknife'} if (defined($self->{'jackknife'})); $jackknife_value++; my $output_id = sprintf("{'protein_tree_id'=>%d, 'clusterset_id'=>1, 'jackknife'=>$jackknife_value}", $self->{'protein_tree'}->node_id); $self->input_job->input_id($output_id); $self->dataflow_output_id($output_id, 2); $self->input_job->update_status('FAILED'); $self->{'protein_tree'}->release_tree; $self->{'protein_tree'} = undef; warn("NJTREE_PHYML : NNI error, resubmitting as jackknife NJTREE_PHYML"); return undef; } } $self->check_job_fail_options; throw("error running njtree phyml, $!\n"); } $self->{'comparaDBA'}->dbc->disconnect_when_inactive(0); } elsif (0 == $self->{'bootstrap'}) { # first part
# ./njtree phyml -nS -f species_tree.nh -p 0.01 -o $BASENAME.cons.nh $BASENAME.nucl.mfa
$cmd = $njtree_phyml_executable; $cmd .= " phyml -nS"; if (defined($self->{'species_tree_file'})) { $cmd .= " -f ". $self->{'species_tree_file'}; } $cmd .= " ". $self->{'input_aln'}; $cmd .= " -p 0.01 "; $self->{'intermediate_newick_file'} = $self->{'input_aln'} . "_intermediate_njtree_phyml_tree.txt "; $cmd .= " -o " . $self->{'intermediate_newick_file'}; $cmd .= " 2>&1 > /dev/null" unless($self->debug); print("$cmd\n") if($self->debug); my $worker_temp_directory = $self->worker_temp_directory; unless(system("cd $worker_temp_directory; $cmd") == 0) { print("$cmd\n"); $self->check_job_fail_options; throw("error running njtree phyml noboot (step 1 of 2), $!\n"); } # second part
# nice -n 19 ./njtree sdi -s species_tree.nh $BASENAME.cons.nh > $BASENAME.cons.nhx
$cmd = $njtree_phyml_executable; $cmd .= " sdi "; if (defined($self->{'species_tree_file'})) { $cmd .= " -s ". $self->{'species_tree_file'}; } $cmd .= " ". $self->{'intermediate_newick_file'}; $cmd .= " 1> " . $self->{'newick_file'}; $cmd .= " 2> /dev/null" unless($self->debug); $self->{'comparaDBA'}->dbc->disconnect_when_inactive(1); print("$cmd\n") if($self->debug); my $worker_temp_directory = $self->worker_temp_directory; unless(system("cd $worker_temp_directory; $cmd") == 0) { print("$cmd\n"); $self->check_job_fail_options; throw("error running njtree phyml noboot (step 2 of 2), $!\n"); } $self->{'comparaDBA'}->dbc->disconnect_when_inactive(0); } else { throw("NJTREE PHYML -- wrong bootstrap option"); } #parse the tree into the datastucture
$self->parse_newick_into_proteintree; my $runtime = time()*1000-$starttime; $self->{'protein_tree'}->store_tag('NJTREE_PHYML_runtime_msec', $runtime);
}
store_proteintreedescriptionprevnextTop
sub store_proteintree {
  my $self = shift;

  return unless($self->{'protein_tree'});

  printf("PHYML::store_proteintree\n") if($self->debug);
  my $treeDBA = $self->{'comparaDBA'}->get_ProteinTreeAdaptor;
  $treeDBA->sync_tree_leftright_index($self->{'protein_tree'});
  $treeDBA->store($self->{'protein_tree'});
  $treeDBA->delete_nodes_not_in_tree($self->{'protein_tree'});

  if($self->debug >1) {
    print("done storing - now print\n");
    $self->{'protein_tree'}->print_tree;
  }

  $self->{'protein_tree'}->store_tag('PHYML_alignment', 'njtree_phyml');
  $self->{'protein_tree'}->store_tag('reconciliation_method', 'njtree_best');
  $self->store_tags($self->{'protein_tree'});

  $self->_store_tree_tags();

  return undef;
}
store_tagsdescriptionprevnextTop
sub store_tags {
  my $self = shift;
  my $node = shift;

  if(defined($self->{'jackknife'})) {
    my $leaf_count = $self->{protein_tree}->num_leaves;
    $self->{protein_tree}->adaptor->delete_tag($self->{protein_tree}->node_id,'gene_count');
    $self->{protein_tree}->store_tag('gene_count', $leaf_count);
  }

  if($node->get_tagvalue("Duplication") eq '1') {
    if($self->debug) { printf("store duplication : "); $node->print_node; }
    $node->store_tag('Duplication', 1);
  } else {
    $node->store_tag('Duplication', 0);
  }

  if (defined($node->get_tagvalue("B"))) {
    my $bootstrap_value = $node->get_tagvalue("B");
    if (defined($bootstrap_value) && $bootstrap_value ne '') {
      if ($self->debug) {
        printf("store bootstrap : $bootstrap_value "); $node->print_node;
      }
      $node->store_tag('Bootstrap', $bootstrap_value);
    }
  }
  if (defined($node->get_tagvalue("DD"))) {
    my $dubious_dup = $node->get_tagvalue("DD");
    if (defined($dubious_dup) && $dubious_dup ne '') {
      if ($self->debug) {
        printf("store dubious_duplication : $dubious_dup "); $node->print_node;
      }
      $node->store_tag('dubious_duplication', $dubious_dup);
    }
  }
  if (defined($node->get_tagvalue("E"))) {
    my $n_lost = $node->get_tagvalue("E");
    $n_lost =~ s/.{2}//;        # get rid of the initial $-
my @lost_taxa = split('-',$n_lost); my %lost_taxa; foreach my $taxon (@lost_taxa) { $lost_taxa{$taxon} = 1; } foreach my $taxon (keys %lost_taxa) { if ($self->debug) { printf("store lost_taxon_id : $taxon "); $node->print_node; } $node->store_tag('lost_taxon_id', $taxon); } } if (defined($node->get_tagvalue("SISi"))) { my $sis_score = $node->get_tagvalue("SISi"); if (defined($sis_score) && $sis_score ne '') { if ($self->debug) { printf("store SISi : $sis_score "); $node->print_node; } $node->store_tag('SISi', $sis_score); } } if (defined($node->get_tagvalue("SISu"))) { my $sis_score = $node->get_tagvalue("SISu"); if (defined($sis_score) && $sis_score ne '') { if ($self->debug) { printf("store SISu : $sis_score "); $node->print_node; } $node->store_tag('SISu', $sis_score); } } if (defined($node->get_tagvalue("SIS"))) { my $sis_score = $node->get_tagvalue("SIS"); if (defined($sis_score) && $sis_score ne '') { if ($self->debug) { printf("store species_intersection_score : $sis_score "); $node->print_node; } $node->store_tag('species_intersection_score', $sis_score); } } if (defined($node->get_tagvalue("SIS1"))) { my $sis_score = $node->get_tagvalue("SIS1"); if (defined($sis_score) && $sis_score ne '') { if ($self->debug) { printf("store SIS1 : $sis_score "); $node->print_node; } $node->store_tag('SIS1', $sis_score); } } if (defined($node->get_tagvalue("SIS2"))) { my $sis_score = $node->get_tagvalue("SIS2"); if (defined($sis_score) && $sis_score ne '') { if ($self->debug) { printf("store SIS2 : $sis_score "); $node->print_node; } $node->store_tag('SIS2', $sis_score); } } # if (defined($node->get_tagvalue("SIS3"))) {
# my $sis_score = $node->get_tagvalue("SIS3");
# if (defined($sis_score) && $sis_score ne '') {
# if ($self->debug) {
# printf("store SIS3 : $sis_score "); $node->print_node;
# }
# $node->store_tag('SIS3', $sis_score);
# }
# }
foreach my $child (@{$node->children}) { $self->store_tags($child); } return undef;
}
write_outputdescriptionprevnextTop
sub write_output {
  my $self = shift;

  $self->check_if_exit_cleanly;
  $self->store_proteintree;
}
General documentation
CONTACTTop
  Contact Albert Vilella on module implementation/design detail: avilella@ebi.ac.uk
Contact Abel Ureta-Vidal on EnsEMBL/Compara: abel@ebi.ac.uk
Contact Ewan Birney on EnsEMBL in general: birney@sanger.ac.uk
APPENDIXTop
The rest of the documentation details each of the object methods.
Internal methods are usually preceded with a _