#!/usr/bin/perl
package Bio::Affymetrix::CHP;
# Docs come before the code
=head1 NAME
Bio::Affymetrix::CHP- parse Affymetrix CHP files
=head1 SYNOPSIS
use Bio::Affymetrix::CHP;
use Bio::Affymetrix::CDF;
# Parse the CDF file
my $cdf=new Bio::Affymetrix::CDF();
$cdf->parse_from_file("foo.cdf");
# Make a new CHP object, using the CDF file
my $chp=new Bio::Affymetrix::CHP($cdf);
# Parse CHP file
$chp->parse_from_file("foo.chp");
# Find some fun facts about this chip
print $chp->algorithm_name()."\n";
print $chp->version()."\n";
# Print out all of the signal values for this chip
while (my ($probename,$results)=each %{$chp->probe_set_results()}) {
print $probename.",".$results->{"Signal"}."\n";
}
=head1 DESCRIPTION
The Affymetrix microarray system produces files in a variety of
formats. If this means nothing to you, these modules are probably not
for you :). This module parses CHP files.
This module requires a Bio::Affymetrix::CDF object before it can do
anything. This must be supplied to the constructor. See the perldoc
for that to see how to use that module. The module can parse various
types of CHP file transparently. You can find out what type you have
by using the original_version() method.
All of the Bio::Affymetrix modules parse a file entirely into
memory. You therefore need enough memory to hold these objects. For
some applications, parsing as a stream may be more appropriate-
hopefully the source to these modules will give enough clues to make
this an easy task.
You can also use this software to write CHP files (see the
write_to_file and write_to_filehandle methods).
=head2 HINTS
You fill the object filled with data using the
parse_from_filehandle, parse_from_string or parse_from_file
routines. You can get/set various statistics using methods on the
object. Data is retrieved as a giant hash from
probe_set_results. Subroutines marked "original_" give values as they
are claimed in the original file, not as they are now (for instance if
you modify the value)
=head1 NOTES
=head2 REFERENCE
Modules were written with the official Affymetrix documentation, which
can be located at http://www.affymetrix.com/support/developer/AffxFileFormats.ZIP
=head2 COMPATIBILITY
This module can parse the CHP files produced by the Affymetrix software
MAS 5 and GCOS v1.2. It can also process files produced by GCOS v1.0
in theory. However the authors of this module have never actually
seen an actual GCOS v1.0 file, and so we rely on the specification
supplied by Affymetrix only. If you have GCOS v1.0 files, feedback
as to whether the code actually works is welcome.
Whatever file format you use the module should work transparently.
These modules are focused on GCOS v1.2 CHP files. The MAS5 CHP
files actually contain a lot of extra information that is not
displayed in MAS5 or GCOS. This information is thrown away by the
parser.
This module can only do expression arrays.
Writing CHP files should work no matter what kind of CHP file was
parsed originally. In other words, you can use these modules as a CHP
file converter.
As stated above, some of the information that is in a MAS5 file is not
present in an GCOS file. This module throws away all of this
additional information. This means that if you try and write a MAS5
file, there will be large blank sections. This should not put off any
programs that use them- this tactic was based on the Affymetrix
conversion.
=head2 KNOWN BUG
Change P Value always comes out wrong when using comparison CHP files
from MAS5. If you have a solution for this, please send email to the
addresses below.
=head1 COPYRIGHT
Copyright (C) 2005 by Nick James, David J Craigon, NASC (arabidopsis.info), The
University of Nottingham
This module is free software. You can copy or redistribute it under the same terms as Perl itself.
Affymetrix is a registered trademark of Affymetrix Inc., Santa
Clara, California, USA.
=head1 AUTHORS
Nick James (nick at arabidopsis.info)
David J Craigon (david at arabidopsis.info)
Nottingham Arabidopsis Stock Centre (http://arabidopsis.info), University of Nottingham.
=head1 METHODS
=cut
use strict;
use warnings;
use Carp;
our $VERSION=0.5;
# Module for processing CHP files
=head2 new
Arg [1] : Bio::Affymetrix::CDF $db_file
Example : my $chp=new Bio::Affymetrix::CHP($cdf);
Description: constructor for CHP object
Returntype : new Bio::Affmetrix::CHP object
Exceptions : none
Caller : general
=cut
sub new {
my $proto = shift;
my $class = ref($proto)||$proto;
my $self = {};
$self->{"cdf"}=shift;
if ((!defined($self->{"cdf"}))||(!$self->{"cdf"}->isa("Bio::Affymetrix::CDF"))) {
croak "Need to supply a Bio::Affymetric::CDF file to Bio::Affymetrix::CHP constructor";
}
bless ($self, $class); # reconsecrate
return $self;
}
# Getter/setter routines
# CDF object associated with this file
=head2 CDF
Arg [1] : Bio::Affymetrix::CDF $db_file
Example : none
Description: get/set for CDF file associated with this CHP file. The db is the one that
belongs to the external_name.
Returntype : Bio::Affymetrix::CDF object
Exceptions : none
Caller : general
=cut
sub CDF {
my $self=shift;
if (my $q=shift) {
$self->{"cdf"}=$q;
}
return $self->{"cdf"};
}
# File format. Either XDA or MAS5 at the moment
=head2 original_format
Arg [0] : none
Example : my $format=$chp->original_format()
Description: Returns the format of the CHP file parsed
(currently) either "XDA" (which is a GCOS v1.2 format,
also known as version 4) or "MAS5" (which is produced
either by MAS 5 or GCOS v1.0, also known as version 3)
Returntype : string ("XDA" or "MAS5")
Exceptions : none
Caller : general
=cut
sub original_format {
my $self=shift;
return $self->{"format"};
}
# Version of file (encoded in file)
=head2 original_version
Arg [0] : none
Example : my $version=$chp->original_version()
Description: Returns the version of the CHP file parsed
Best used in conjunction with original_format,
above. For XDA files, the version is always
(currently) 1. For MAS5 files the version is either 12
for a file produced by MAS5, or 13 for a GCOS v1.0
file. Code is written for parsing version 13 files,
but we\'ve no idea if it works.
Returntype : string
Exceptions : none
Caller : general
=cut
sub original_version {
my $self=shift;
return $self->{"version"};
}
# Number of columns in the array
=head2 cols
Arg [0] : none
Example : my $x=$chp->cols()
Description: Numbers of columns in the array
Returntype : integer
Exceptions : none
Caller : general
=cut
sub cols {
my $self=shift;
if (my $q=shift) {
$self->{"no_cols"}=$q;
}
return $self->{"no_cols"};
}
# Number of rows in the array
=head2 rows
Arg [0] : none
Example : my $y=$chp->rows()
Description: Get/set numbers of rows in the array
Returntype : integer
Exceptions : none
Caller : general
=cut
sub rows {
my $self=shift;
if (my $q=shift) {
$self->{"no_rows"}=$q;
}
return $self->{"no_rows"};
}
# Number of probes claimed
=head2 original_number_of_probes
Arg [0] : none
Example : my $original_probes=$chp->original_number_of_probes()
Description: Gets the original number of probes reported in the
array.
The CHP files have the number of probes stored in
them, and this function lets you read this number as
it was stored in the file originally.
A better way of finding out the current number of
probes is to count the number in the probe_set_results
hash, like so: scalar(keys %{$chp->probe_set_results()});
Returntype : integer
Exceptions : none
Caller : general
=cut
sub original_number_of_probes {
my $self=shift;
return $self->{"no_units"};
}
# Number of qc units
=head2 original_number_qc_units
Arg [0] : none
Example : my $original_qc=$chp->original_number_qc_units()
Description: Gets the original number of QC units in the file.
Returntype : integer
Exceptions : none
Caller : general
=cut
sub original_number_qc_units {
my $self=shift;
return $self->{"no_qc_units"};
}
# MS COM prog ID
=head2 original_com_progid
Arg [0] : none
Example : my $com_id=$chp->original_com_progid()
Description: Gets the progid of the original Microsoft COM object that made
this CHP file
Returntype : string
Exceptions : none
Caller : general
=cut
sub original_com_progid {
my $self=shift;
return $self->{"com_progid"};
}
# CEL file name this CHP file originated from
=head2 CEL_file_name
Arg [1] : string $cel_file_name (optional)
Example : my $cel_file_name=$chp->CEL_file_name();
Description: Get/set the CEL file this CHP file was made from
Returntype : string
Exceptions : none
Caller : general
=cut
sub CEL_file_name {
my $self=shift;
if (my $q=shift) {
$self->{"cel_file_name"}=$q;
}
return $self->{"cel_file_name"};
}
=head2 original_file_name
Arg [0] : none
Example : my $chp_file_name=$chp->original_file_name();
Description: If this object was created using parse_from_file, the original filename. Otherwise undef.
Returntype : string
Exceptions : none
Caller : general
=cut
sub original_file_name {
my $self=shift;
return $self->{"file_name"};
}
# ATH1, etc.
=head2 probe_array_type
Arg [1] : string $array_type (optional)
Example : my $probe_array_type=$chp->probe_array_type();
Description: Get/set the Affymetrix chip type used in the
production of this CHP file. String is same format as CDF file name,
for example ATH1-121501
Returntype : string
Exceptions : none
Caller : general
=cut
sub probe_array_type {
my $self=shift;
if (my $q=shift) {
$self->{"probe_array_type"}=$q;
}
return $self->{"probe_array_type"};
}
# Algorithm name
=head2 algorithm_name
Arg [1] : string $algorithm_name (optional)
Example : my $algorithm_name=$chp->algorithm_name();
Description: Get/set the algorithm name that turned the CEL file
into this CHP file
Returntype : string
Exceptions : none
Caller : general
=cut
sub algorithm_name {
my $self=shift;
if (my $q=shift) {
$self->{"algorithm_name"}=$q;
}
return $self->{"algorithm_name"};
}
=head2 algorithm_version
Arg [1] : string $algorithm_version (optional)
Example : my $algorithm_version=$chp->algorithm_version();
Description: Get/set the algorithm version that turned the CEL file
into this CHP file
Returntype : string
Exceptions : none
Caller : general
=cut
sub algorithm_version {
my $self=shift;
if (my $q=shift) {
$self->{"algorithm_version"}=$q;
}
return $self->{"algorithm_version"};
}
# Algorithm params- returns hashref
=head2 algorithm_params
Arg [1] : hashref $algorithm_params (optional)
Example : my %algorithm_params=%{$chp->algorithm_params()};
# Print scale factor
print $algorithm_params("SF");
Description: Get/set the algorithm parameters for turning the CEL
file into this CHP file. Returns a reference to a hash, keyed on
parameter name.
Parameters include:
SF (scale factor)
Alpha1
Alpha2
Tau
NF (normalisation factor)
Returntype : hashref
Exceptions : none
Caller : general
=cut
sub algorithm_params {
my $self=shift;
if (my $q=shift) {
$self->{"algorithm_params"}=$q;
}
return $self->{"algorithm_params"};
}
# Summary Statistics- returns hashref
=head2 summary_statistics
Arg [1] : hashref $summary_statistics (optional)
Example : my %summary_statistics=%{$chp->summary_statistics()};
# Print RawQ
print $chp->summary_statistics()->{"RawQ"};
Description: Get/set the summary statistics for turning the CEL
file into this CHP file. Returns a reference to a hash, keyed on
parameter name.
Parameters include:
RawQ
Noise
Background
Returntype : hashref
Exceptions : none
Caller : general
=cut
sub summary_statistics {
my $self=shift;
if (my $q=shift) {
$self->{"summary_statistics"}=$q;
}
return $self->{"summary_statistics"};
}
# Used in calculating background
=head2 smooth_factor
Arg [1] : float $smooth_factor (optional)
Example : my $smooth_factor=$chp->smooth_factor();
Description: Get/set the smooth factor. Your guess is as good as
mine as to what this actually is, although it is used in calculating
the background. Not available in MAS5 files.
Returntype : float
Exceptions : none
Caller : general
=cut
sub smooth_factor {
my $self=shift;
if (my $q=shift) {
$self->{"smooth_factor"}=$q;
}
return $self->{"smooth_factor"};
}
# An arrayref of zones
=head2 background_zones
Arg [1] : arrayref of arrayrefs $background_zones (optional)
Example : my @background_zones=@{$chp->background_zones()};
# Print "X", "Y", "Background Value" for background zone 0
print $background_zones[0]->[0],$background_zones[0]->[1],$background_zones[0]->[2];
Description: Get/set an array of background zones. Again, your guess is as good as
mine as to what this actually is, although it is used in calculating
the background. Not available in MAS5 files.
Returns an arrayref to an arrayref. Each zone has an array three
values long, which are X,Y and background value respectively.
Returntype : arrayref of arrayrefs
Exceptions : none
Caller : general
=cut
sub background_zones {
my $self=shift;
if (my $q=shift) {
$self->{"zones"}=$q;
}
return $self->{"zones"};
}
# The hash of results
=head2 probe_set_results
Arg [1] : hashref of hashrefs $probe_set_results (optional)
Example : my %results=%{$chp->probe_set_results()};
e
# Print "Signal", "Detection Call", "StatPairsUsed" for my favourite probe
print ($results{"246310_at"}->{"Signal"}."\n".$results{"246310_at"}->{"DetectionCall"}."\n".$results{"246310_at"}->{"StatPairsUsed"}."\n");
Description: Gain access to the actual data. Returns a reference to
hash, keyed on probe name. Each value contains another reference
to a hash with the following keys:
Signal
DetectionCall (detection call- one of P M A N)
DetectionPValue
StatPairs
StatPairsUsed
Probeset (a Bio::CDF::ProbeSet object, q.v.)
and optionally:
Change
ChangePValue
SignalLogRatio
SignalLogRatioHigh
SignalLogRatioLow
CommonPairs
when it is a CHP files with a comparison in it.
Returntype : hashref of hashrefs
Exceptions : none
Caller : general
=cut
sub probe_set_results {
my $self=shift;
if (my $q=shift) {
$self->{"probe_set_results"}=$q;
}
return $self->{"probe_set_results"};
}
# Parsing
=head2 parse_from_string
Arg [1] : string
Example : $chp->parse_from_string($chp_file_in_a_string);
Description: Parse a CHP file from a buffer in memory
Returntype : none
Exceptions : none
Caller : general
=cut
sub parse_from_string {
my $self=shift;
my $string=shift;
open CHP,"<",\$string or croak "Cannot open string stream";
$self->parse_from_filehandle(\*CHP);
close CHP;
}
=head2 parse_from_file
Arg [1] : string
Example : $chp->parse_from_file($chp_filename);
Description: Parse a CHP file from a file
Returntype : none
Exceptions : dies if can\'t open file
Caller : general
=cut
sub parse_from_file {
my $self=shift;
my $filename=shift;
open CHP,"<".$filename or croak "Cannot open file ".$filename;
$self->{"file_name"}=$filename;
$self->parse_from_filehandle(\*CHP);
close CHP;
}
=head2 parse_from_filehandle
Arg [1] : reference to filehandle
Example : $chp->parse_from_filehandle(\*STDIN);
Description: Parse a CHP file from a filehandle
Returntype : none
Exceptions : none
Caller : general
=cut
sub parse_from_filehandle {
my $self=shift;
my $fh=shift;
binmode $fh;
# First step- detect whether it's a GCOS or MAS5 file
# A buffer for reading things into
my $buffer;
# XDA files have their first feature as a magic number, 65
read ($fh, $buffer, 4);
my $magic_number = unpack("V", $buffer);
if ($magic_number==65) {
$self->_parse_xda($fh);
return;
}
# MAS5 files have their first feature as a string
read ($fh, $buffer, 18, 4);
$magic_number=unpack("A22",$buffer);
if ($magic_number eq "GeneChip Sequence File") {
# It's a MAS5/GCOS v1.0 "v3 file"! Yippee!
$self->_parse_mas5($fh);
return;
}
croak "This doesn't look like a CHP file to me. I can only understand certain CHP filetypes, however\n";
}
sub _parse_xda {
my ($self,$fh) = @_;
$self->{"format"}="XDA";
my $buffer;
# First some trivia
read ($fh, $buffer, 4);
$self->{"version"}= unpack ("V", $buffer);
if ($self->{"version"}!=1) {
carp "This CHP file is newer than the software parsing them. Results may be suspect."; # die here, perhaps?
}
read ($fh, $buffer, 12);
($self->{"no_cols"},$self->{"no_rows"},$self->{"no_units"},$self->{"no_qc_units"})= unpack ("S2V2", $buffer);
read ($fh, $buffer, 4);
$self->{"chip_type"}=unpack ("V", $buffer);
if ($self->{"chip_type"}!=0) {
croak "This software does not process non-expression arrays";
}
$self->{"com_progid"}=$self->unpack_length_string($fh);
$self->{"cel_file_name"}=$self->unpack_length_string($fh);
$self->{"probe_array_type"}=$self->unpack_length_string($fh);
$self->{"algorithm_name"}=$self->unpack_length_string($fh);
$self->{"algorithm_version"}=$self->unpack_length_string($fh);
# Algorithm parameters
{
read ($fh, $buffer, 4);
my $no_algorithm_params = unpack ("V", $buffer);
# get varying number of parameter names and values and read into hash
my %algorithm_params;
for (my $i=0;$i<$no_algorithm_params;$i++) {
my $name=$self->unpack_length_string($fh);
my $value=$self->unpack_length_string($fh);
$algorithm_params{$name}=$value;
}
$self->{"algorithm_params"}=\%algorithm_params;
}
# Summary statistics
{
read ($fh, $buffer, 4);
my $no_stats = unpack ("V", $buffer);
# get varying number of parameter names and values and read into hash
my %h;
for (my $i=0;$i<$no_stats;$i++) {
my $name=$self->unpack_length_string($fh);
my $value=$self->unpack_length_string($fh);
$h{$name}=$value;
}
$self->{"summary_statistics"}=\%h;
}
# Background calculation
{
read ($fh, $buffer, 8);
my $m;
($m,$self->{"smooth_factor"}) = unpack ("Vf", $buffer);
$self->{"zones"}=[];
my %zone_info;
for (my $i=0;$i<$m;$i++) {
read ($fh, $buffer, 12);
my @zone = unpack ("f3", $buffer);
push @{$self->{"zones"}},\@zone;
}
}
# Actual data. This is the bit that would need to be added to, if we did SNP etc. arrays
{
my $size;
read ($fh, $buffer, 5);
($self->{"analysis_type"},$size)=unpack("CV", $buffer);
my %data;
my $probesetlist=$self->{"cdf"}->probesets();
foreach my $i (sort {int($a)<=>int($b)} keys(%$probesetlist)) {
read ($fh, $buffer, $size);
# Non-comparison analysis
if ($self->{"analysis_type"}==0 || $self->{"analysis_type"}==2) {
my @results=unpack("Cf2S2",$buffer);
my %h;
if ($results[0]==0) {
$h{"DetectionCall"}="P";
} elsif ($results[0]==1) {
$h{"DetectionCall"}="M";
} elsif ($results[0]==2) {
$h{"DetectionCall"}="A";
} elsif ($results[0]==3) {
$h{"DetectionCall"}="N";
}
$h{"DetectionPValue"}=$results[1];
$h{"Signal"}=$results[2];
$h{"StatPairs"}=$results[3];
$h{"StatPairsUsed"}=$results[4];
$h{"Probeset"}=$probesetlist->{$i};
$data{$probesetlist->{$i}->name()}=\%h;
} else {
# Comparison analysis
$self->{"comparison"}=1;
my @results=unpack("Cf2S2Cf4S",$buffer);
my %h;
if ($results[0]==0) {
$h{"DetectionCall"}="P";
} elsif ($results[0]==1) {
$h{"DetectionCall"}="M";
} elsif ($results[0]==2) {
$h{"DetectionCall"}="A";
} elsif ($results[0]==3) {
$h{"DetectionCall"}="N";
}
$h{"DetectionPValue"}=$results[1];
$h{"Signal"}=$results[2];
$h{"StatPairs"}=$results[3];
$h{"StatPairsUsed"}=$results[4];
$h{"Change"}=$results[6];
$h{"ChangePValue"}=$results[7];
$h{"SignalLogRatio"}=$results[8];
$h{"SignalLogRatioLow"}=$results[9];
$h{"SignalLogRatioHigh"}=$results[10];
$h{"CommonPairs"}=$results[11];
$h{"Probeset"}=$probesetlist->[$i];
$data{$probesetlist->{$i}->name()}=\%h;
}
}
$self->{"probe_set_results"}=\%data;
}
}
sub _parse_mas5 {
my ($self,$fh) = @_;
$self->{"format"}="MAS5";
my $buffer;
# Get version from file
read ($fh, $buffer, 4);
$self->{"version"}=unpack ("V", $buffer);
if ($self->{"version"}!=12&&$self->{"version"}!=13) {
carp "This CHP file has a version number unrecognised by the software parsing them. Results may be suspect."; # die here, perhaps?
}
if ($self->{"version"}==13) {
carp "The authors of this module have never seen a genuine GCOS v1.0 CHP file. This program can parse them, but we're only relying on the specification supplied by Affymetrix- we've not tested this at all. Suspect results therefore are highly likely.";
}
# Trivia section
$self->{"algorithm_name"}=$self->unpack_length_string($fh);
$self->{"algorithm_version"}=$self->unpack_length_string($fh);
# Parse algorithm parameters to maintain compatability with XDA format
{
my %algorithm_params;
my $parse_me=$self->unpack_length_string($fh);
foreach my $i (split / /,$parse_me) {
my ($name,$value)=split /=/,$i;
$algorithm_params{$name}=$value;
}
$self->{"algorithm_params"}=\%algorithm_params;
}
# Summary statistics
{
my %summary_stats;
my $parse_me=$self->unpack_length_string($fh);
foreach my $i (split / /,$parse_me) {
my ($name,$value)=split /=/,$i;
$summary_stats{$name}=$value;
}
$self->{"summary_statistics"}=\%summary_stats;
}
read ($fh, $buffer, 20);
my $max_probeset_num;
($self->{"no_rows"},$self->{"no_cols"},$self->{"no_units"},$max_probeset_num,$self->{"no_qc_units"})= unpack ("V5", $buffer);
# THROW AWAY PROBESET NUMBER FOR EACH PROBESET
read ($fh, $buffer, 4*$self->{"no_units"});
# THROW AWAY NUMBER OF PROBE PAIRS FOR EACH PROBESET
read ($fh, $buffer, 4*$max_probeset_num);
# THROW AWAY PROBESETTYPE FOR EACH PROBESET
read ($fh, $buffer, 4*$max_probeset_num); # Should test
# THROW AWAY PROBESET NUMBER FOR EACH PROBESET
read ($fh, $buffer, 4*$self->{"no_units"});
read ($fh, $buffer, 512);
($self->{"probe_array_type"},$self->{"cel_file_name"})=unpack ("Z256Z256",$buffer);
if ($self->{"probe_array_type"} ne $self->{"cdf"}->name()) {
carp "The CDF object you have supplied does not have the same name as the CDF file used to make this CHP file. Results may be dubious";
}
$self->{"com_progid"}=$self->unpack_length_string($fh);
# Actual data. This is the bit that would need to be added to, if we did SNP etc. arrays
{
my %data;
if ($self->{"version"}==12) {
my %results;
my $probesetlist=$self->{"cdf"}->probesets();
foreach my $i (sort {int($a)<=>int($b)} keys(%$probesetlist)) {
# Non-comparison analysis
read ($fh, $buffer, 44);
my @results=unpack("V7f3V", $buffer);
my %h;
if ($results[10]==0) {
$h{"DetectionCall"}="P";
} elsif ($results[10]==1) {
$h{"DetectionCall"}="M";
} elsif ($results[10]==2) {
$h{"DetectionCall"}="A";
} elsif ($results[10]==3) {
$h{"DetectionCall"}="N";
}
$h{"DetectionPValue"}=$results[7];
$h{"Signal"}=$results[9];
$h{"StatPairs"}=$results[0];
$h{"StatPairsUsed"}=$results[1];
$h{"Probeset"}=$probesetlist->{$i};
# Blimey-o-reilly! There's an entire CEL file in here! Let's ditch that.
read ($fh, $buffer, $h{"StatPairs"}*52); #52 bytes of junk per probeset?
read ($fh, $buffer, 4);
# Comparison analysis
if (unpack("V",$buffer)==1) {
$self->{"comparison"}=1;
read ($fh, $buffer, 58);
my @results=unpack("V5c2l9",$buffer);
$h{"Change"}=$results[4];
$h{"ChangePValue"}=$results[15]/1000;
$h{"SignalLogRatio"}=$results[12]/1000;
$h{"SignalLogRatioLow"}=$results[14]/1000;
$h{"SignalLogRatioHigh"}=$results[9]/1000;
$h{"CommonPairs"}=$results[0];
$h{"BaselineAbsent"}=$results[5];
$h{"Probeset"}=$probesetlist->{$i};
}
# $probesetlist->[$i] or die "Suspect CDF file! We have more data in the CHP file than expected. Are you using the right CDF file?";
$data{$probesetlist->{$i}->name()}=\%h;
}
} elsif ($self->{"version"}==13) {
my %results;
my $probesetlist=$self->{"cdf"}->probesets();
for (my $i=0;$i<$self->{"no_units"};$i++) {
# Non-comparison analysis
read ($fh, $buffer, 24);
my @results=unpack("V3f2V", $buffer);
my %h;
if ($results[5]==0) {
$h{"DetectionCall"}="P";
} elsif ($results[5]==1) {
$h{"DetectionCall"}="M";
} elsif ($results[5]==2) {
$h{"DetectionCall"}="A";
} elsif ($results[5]==3) {
$h{"DetectionCall"}="N";
}
$h{"DetectionPValue"}=$results[3];
$h{"Signal"}=$results[4];
$h{"StatPairs"}=$results[0];
$h{"StatPairsUsed"}=$results[1];
$h{"Probeset"}=$probesetlist->[$i];
# Blimey-o-reilly! There's still-enough-of-a-CEL-file in here to be annoying! Nurse! The screens!
read ($fh, $buffer, $h{"StatPairs"}*20); #20 bytes of junk per probeset?
read ($fh, $buffer, 4);
# Comparison analysis
if (unpack("V",$buffer)==1) {
read ($fh, $buffer, 27); #????????
my @results=unpack("V2cV4",$buffer);
my %h;
$h{"Change"}=$results[1];
$h{"ChangePValue"}=$results[6];
$h{"SignalLogRatio"}=$results[3]/1000;
$h{"SignalLogRatioLow"}=$results[4]/1000;
$h{"SignalLogRatioHigh"}=$results[5]/1000;
$h{"CommonPairs"}=$results[0];
$h{"BaselineAbsent"}=$results[2];
$h{"Probeset"}=$probesetlist->[$i];
}
$probesetlist->[$i] or die "Suspect CDF file! We have more data in the CHP file than expected. Are you using the right CDF file?";
$data{$probesetlist->[$i]->name()}=\%h;
}
}
$self->{"probe_set_results"}=\%data;
}
}
# binary file utility to read a length-defined string
sub unpack_length_string {
my $self=shift;
my $fh=shift;
my $buffer;
read ($fh, $buffer, 4);
my $len = unpack ("V", $buffer);
read ($fh, $buffer, $len);
return unpack("a".$len,$buffer);
}
=head2 write_to_file
Arg [1] : string $filename
Arg [2] : string $format
Arg [3] : string $version
Example : $cdf->write_to_file($cdf_filename);
Description: Writes a CDF file to a file. See write_to_filehandle for descriptions of format and version
Returntype : none
Exceptions : dies if cannot open file
Caller : general
=cut
sub write_to_file {
my $self=shift;
my $filename=shift;
open CDF,">".$filename or die "Cannot open file for writing".$filename;
$self->write_to_filehandle(\*CDF,@_);
close CDF;
}
=head2 write_to_filehandle
Arg [1] : filehandle $filehandle
Arg [2] : string $format
Arg [3] : string $version
Example : $chp->write_to_filehandle(\*STDOUT);
Description: Writes a CDF file to a filehandle. Takes arguments of
the filehandle, the desired format, and the desired version of that
format.
Currently, format defaults to MAS5, and version defaults to
GC3.0.
Returntype : none
Exceptions : dies if cannot open file
Caller : general
=cut
sub write_to_filehandle {
my $self=shift;
my $filehandle=shift;
my $format=shift;
my $version=shift;
my $fake=shift;
if (!defined $format) {
$format="MAS5";
$version=12;
}
if ($format eq "XDA" && !defined $version) {
$version=1;
}
if ($format eq "XDA") {
$self->_write_xda($filehandle,$version,$fake);
} elsif ($format eq "MAS5") {
$self->_write_mas5($filehandle,$version,$fake);
} else {
croak "Format must be XDA or MAS5";
}
}
sub _write_xda {
my $self=shift;
my $filehandle=shift;
my $version=shift;
my $fake=shift;
if ($version!=1) {
croak "Bio::Affymetrix can only write version 1 XDA files";
}
# Static stuff- always the same
print $filehandle pack ("V2S2V3",65,$version,$self->{"no_cols"},$self->{"no_rows"},$self->{"no_units"},$self->{"no_qc_units"},0); # 0 at the end means expression array. Check no_units, no_qc_units!
if (!$fake) {
print $filehandle pack ("V/a*","GeneChip.CallGEBaseCall.1");
} else {
print $filehandle pack ("V/a*","Bio::Affymetrix version ".$VERSION);
}
print $filehandle pack ("V/a*",$self->{"cel_file_name"});
print $filehandle pack ("V/a*",$self->{"probe_array_type"});
print $filehandle pack ("V/a*",$self->{"algorithm_name"});
print $filehandle pack ("V/a*",$self->{"algorithm_version"});
print $filehandle pack ("V",scalar(keys %{$self->{"algorithm_params"}}));
while (my ($key,$value)=each %{$self->{"algorithm_params"}}) {
print $filehandle pack ("V/a*V/a*",$key,$value);
}
print $filehandle pack ("V",scalar(keys %{$self->{"summary_statistics"}}));
while (my ($key,$value)=each %{$self->{"summary_statistics"}}) {
print $filehandle pack ("V/a*V/a*",$key,$value);
}
print $filehandle pack ("Vf",scalar(@{$self->{"zones"}}),$self->{"smooth_factor"});
foreach my $i (@{$self->{"zones"}}) {
print $filehandle pack ("f3",@$i);
}
print $filehandle pack("C", $self->{"analysis_type"});
# Hand calculated size of objects
if ($self->{"analysis_type"}==0 || $self->{"analysis_type"}==2) {
print $filehandle pack("V", 13);
} else {
print $filehandle pack("V", 32);
}
# Resequencing - BLANKED
my $probesetlist=$self->{"cdf"}->probesets();
foreach my $i (sort {int($a)<=>int($b)} keys(%$probesetlist)) {
if ($self->{"analysis_type"}==0 || $self->{"analysis_type"}==2) {
my $result=$self->{"probe_set_results"}->{$probesetlist->{$i}->name()};
my $dt;
if ($result->{"DetectionCall"} eq "P") {
$dt=0;
} elsif ($result->{"DetectionCall"} eq "M") {
$dt=1;
} elsif ($result->{"DetectionCall"} eq "A") {
$dt=2;
} elsif ($result->{"DetectionCall"} eq "N") {
$dt=3;
}
print $filehandle pack("Cf2S2",$dt,$result->{"DetectionPValue"},$result->{"Signal"},$result->{"StatPairs"},$result->{"StatPairsUsed"});
} else {
# Comparison analysis
my $result=$self->{"probe_set_results"}->{$probesetlist->{$i}->name()};
my $dt;
if ($result->{"DetectionCall"} eq "P") {
$dt=0;
} elsif ($result->{"DetectionCall"} eq "M") {
$dt=1;
} elsif ($result->{"DetectionCall"} eq "A") {
$dt=2;
} elsif ($result->{"DetectionCall"} eq "N") {
$dt=3;
}
print $filehandle pack("Cf2S2Cf4S",$dt,$result->{"DetectionPValue"},$result->{"Signal"},$result->{"StatPairs"},$result->{"StatPairsUsed"},$result->{"Change"},$result->{"ChangePValue"},$result->{"SignalLogRatio"},$result->{"SignalLogRatioLow"},$result->{"SignalLogRatioHigh"},$result->{"CommonPairs"});
}
}
}
sub _write_mas5 {
my $self=shift;
my $filehandle=shift;
my $version=shift;
my $fake=shift;
if (!defined $version) {
$version=12;
}
if ($version!=12) {
croak "Unfortuantely, these modules can only write version 12 MAS5 files at present. If you've got software that can make or read version 13 files, please write to the authors of Bio::Affymetrix";
}
print $filehandle pack("a22VV/a*V/a*","GeneChip Sequence File",$version,$self->{"algorithm_name"},$self->{"algorithm_version"});
{
# Make parameters and summary stats
my $algoparams;
while (my ($key,$value)=each %{$self->{"algorithm_params"}}) {
$algoparams.=$key."=".$value." ";
}
my $algosummary;
while (my ($key,$value)=each %{$self->{"summary_statistics"}}) {
$algosummary.=$key."=".$value." ";
}
print $filehandle pack("V/a*V/a*",$algoparams,$algosummary);
}
# Rows,columns come from CDF file
print $filehandle pack("V2",(scalar(@{$self->CDF()->probe_grid()})+1),(scalar(@{$self->CDF()->probe_grid()->[1]})+1)); ##Maybe?
my @probelist=sort {int($a)<=>int($b)} (keys %{$self->CDF()->probesets()});
# Number of probes trivia
print $filehandle pack("V3",scalar(@probelist),$probelist[scalar(@probelist)-1],$self->{"no_qc_units"});
print $filehandle pack("V*",@probelist);
# This bit is surely a mistake. They want the number of probe pairs per probeset, until probeset number is greater than the number of probesets
foreach my $i (@probelist) {
if ($i<scalar(@probelist)) {
print $filehandle pack("V",scalar(@{$self->CDF()->probesets()->{$i}->probe_pairs()}));
} else {
print $filehandle pack("V",0);
}
}
foreach my $i (@probelist) {
print $filehandle pack("V",(3));
}
foreach my $i (@probelist) {
print $filehandle pack("V",scalar(@{$self->CDF()->probesets()->{$i}->probe_pairs()->[0]}));
}
print $filehandle pack("a256a256",$self->{"probe_array_type"},$self->{"cel_file_name"});
if (!$fake) {
print $filehandle pack ("V/a*","GeneChip.CallGEBaseCall.1");
} else {
print $filehandle pack ("V/a*","Bio::Affymetrix version ".$VERSION);
}
}
1;