/usr/bin/mipeCheckSanity is in mipe 1.1-6.
This file is owned by root:root, with mode 0o755.
The actual contents of the file can be viewed below.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 | #!/usr/bin/perl
# This library is free software; you can redistribute it and/or
# modify it under the terms of the GNU Lesser General Public
# License as published by the Free Software Foundation; either
# version 2.1 of the License, or (at your option) any later version.
#
# This library is distributed in the hope that it will be useful,
# but WITHOUT ANY WARRANTY; without even the implied warranty of
# MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU
# Lesser General Public License for more details.
#
# You should have received a copy of the GNU Lesser General Public
# License along with this library ('COPYING'); if not, write to the Free Software
# Foundation, Inc., 51 Franklin St, Fifth Floor, Boston, MA 02110-1301 USA
use strict;
#use warnings;
use XML::Twig;
use Data::Dumper;
=head1 NAME
mipeCheckSanity.pl - Check sanity of MIPE file, focussing on contents (vs 'form', that can be checked using xmllint)
based on MIPE version v0.9
arguments: * mipe_file
* (optional) list of PCR IDs
=head1 SYNOPSIS
mipe2genotypes.pl your_file.mipe <pcr_id1> <pcr_id2>
=head1 ADDITIONAL INFO
See http://mipe.sourceforge.net
=head1 AUTHOR
Jan Aerts (jan.aerts@bbsrc.ac.uk)
=cut
my %amb_codes = ( M => ['A', 'C']
, R => ['A', 'G']
, W => ['A', 'T']
, S => ['C', 'G']
, Y => ['C', 'T']
, K => ['G', 'T']
, V => ['A', 'C', 'G']
, H => ['A', 'C', 'T']
, D => ['A', 'G', 'T']
, B => ['C', 'G', 'T']
, N => ['A', 'C', 'G', 'T']
);
my ( $file, @pcr_ids ) = @ARGV;
if ( not defined $file ) { die "Please provide filename\n" };
my $twig = XML::Twig->new( TwigHandlers => { pcr => \&pcr }
, pretty_print => 'indented' );
$twig->parsefile($file);
exit;
sub pcr {
my ( $twig, $pcr ) = @_;
my $to_include = 0;
my $pcr_id = $pcr->first_child('id')->text;
if ( scalar @pcr_ids > 0 ) {
$to_include = 0;
foreach ( @pcr_ids ) {
if ( $pcr_id =~ /$_/i ) {
$to_include = 1;
}
}
} else {
$to_include = 1;
}
if ( $to_include ) {
#####Check that there is a USE part
if ( not defined $pcr->first_child('use') ) {
print "WARNING\tCHECK01\t", $pcr->first_child('id')->text, "\tNo use part defined\n";
next;
}
#####Check that snp_id in genotypes actually is defined as a SNP
{
my @snps = $pcr->first_child('use')->children('snp');
my %snps;
foreach my $snp ( @snps ) {
$snps{$snp->first_child('id')->text} = 1;
}
my @samples = $pcr->first_child('use')->children('sample');
foreach my $sample ( @samples ) {
my @sample_snps = $sample->children('genotype');
foreach my $sample_snp ( @sample_snps ) {
if ( not defined $snps{$sample_snp->first_child('snp_id')->text} ) {
print "ERROR\tCHECK02\t", $pcr_id, "\tSample ", $sample->first_child('id')->text, ": genotype SNP ID: ", $sample_snp->first_child('snp_id')->text, " not defined as SNP\n";
}
}
}
}
#####Check if SBE primer sequence and SNP position are in concordance with piece of use seq
{
my $use_seq = $pcr->first_child('use')->first_child('seq')->text;
my @snps = $pcr->first_child('use')->children('snp');
foreach my $snp ( @snps ) {
my $snp_pos = $snp->first_child('pos')->text;
my @assays = $snp->children('assay');
foreach my $assay ( @assays ) {
if ( (uc $assay->first_child('type')->text) eq 'SBE' ) {
my $sbe_specific = $assay->first_child('specific')->text;
my $sbe_strand = $assay->first_child('strand')->text;
if ( uc $sbe_strand =~ /^F/ ) {
if ( not uc $sbe_specific eq uc substr($use_seq, ($snp_pos-(length $sbe_specific)-1), length $sbe_specific) ) {
print "ERROR\tCHECK03\t", $pcr->first_child('id')->text, "\tSBE_SEQ does not comply with SNP pos and USE SEQ\t", $assay->first_child('id')->text, "\tSBE=", $sbe_specific, "\tUSE=", substr($use_seq, ($snp_pos-(length $sbe_specific)), length $sbe_specific), "\n";
}
} else {
$sbe_specific = reverse $sbe_specific;
$sbe_specific =~ tr/ACGTacgt/TGCAtgca/;
if ( not uc $sbe_specific eq uc substr($use_seq, $snp_pos, length $sbe_specific) ) {
print "ERROR\tCHECK03\t", $pcr->first_child('id')->text, "\tSBE_SEQ does not comply with SNP pos and USE SEQ\t", $assay->first_child('id')->text, "\tSBE=", $sbe_specific, "\tUSE=", substr($use_seq, $snp_pos, length $sbe_specific), "\n";
}
}
}
}
}
}
#####Check if PCR primers can be found back in DESIGN sequence
{
my $design = $pcr->first_child('design');
my $primer1_seq = uc $design->first_child('primer1')->first_child('seq')->text;
( my $primer2_seq = reverse uc $design->first_child('primer2')->first_child('seq')->text ) =~ tr/ACGTacgt/TGCAtgca/;
my $design_seq = uc $design->first_child('seq')->text;
if ( not $design_seq =~ /$primer1_seq/ ) {
print "ERROR\tCHECK04\t", $pcr->first_child('id')->text, "\tPRIMER1_SEQ cannot be found back in DESIGN_SEQ\t", $primer1_seq, "\n";
}
if ( not $design_seq =~ /$primer2_seq/ ) {
print "ERROR\tCHECK04\t", $pcr->first_child('id')->text, "\tPRIMER2_SEQ cannot be found back in DESIGN_SEQ\t", $primer2_seq, "\n";
}
}
#####Check if SNP ambiguity code is in concordance with USE sequence
{
my $use_seq = $pcr->first_child('use')->first_child('seq')->text;
my @snps = $pcr->first_child('use')->children('snp');
foreach my $snp ( @snps ) {
if ( not defined $snp->first_child('amb') ) { next };
my $snp_id = $snp->first_child('id')->text;
my $snp_pos = $snp->first_child('pos')->text;
my $snp_amb = $snp->first_child('amb')->text;
my $use_allele = substr($use_seq, $snp_pos-1, 1);
my $good = 0;
if ( uc $use_allele eq uc $snp_amb ) {
$good = 1;
}
if ( uc $use_allele eq 'N' ) {
$good = 1;
}
foreach my $amb_allele ( @{$amb_codes{$snp_amb}} ) {
if ( uc $amb_allele eq uc $use_allele ) {
$good = 1;
}
}
if ( not $good ) {
print "ERROR\tCHECK05\t", $pcr->first_child('id')->text, "\t", $snp->first_child('id')->text, "\tSNP ambiguity code (=", $snp_amb, ") does not comply with allele at SNP position in USE seq (=", $use_allele, ") (check CHECK03)\n";
}
}
}
#####Check for empty elements
{
if ( $pcr->first_child('id')->text eq '' ) {
print "ERROR\tCHECK06\tNo PCR ID\n";
}
my @descendants = $pcr->descendants;
foreach my $descendant ( @descendants ) {
if ( scalar $descendant->descendants == 0 ) {
if ( $descendant->text eq '' ) {
print "WARNING\tCHECK06\tEmpty element in ", $pcr->first_child('id')->text, "\n";
}
}
}
}
}
}
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