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# POD documentation - main docs before the code |
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3
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=head1 NAME |
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5
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GenOO::Data::File::SAM::Cigar - Role that corresponds to the SAM file CIGAR string |
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7
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=head1 SYNOPSIS |
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This role when consumed requires specific attributes and provides |
10
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methods to extract information from the CIGAR string as defined in |
11
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the SAM format specifications. |
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13
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=head1 DESCRIPTION |
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15
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The cigar string describes the alignment of a query on a reference sequence. |
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This role offers methods that can extract information from the CIGAR string |
17
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directly such as the positions on insertions, the total number of deletions, etc |
18
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19
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The CIGAR operations are given in the following table (set `*' if unavailable): |
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Op BAM Description |
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M 0 alignment match (can be a sequence match or mismatch) |
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I 1 insertion to the reference |
23
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D 2 deletion from the reference |
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N 3 skipped region from the reference |
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S 4 soft clipping (clipped sequences present in SEQ) |
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H 5 hard clipping (clipped sequences NOT present in SEQ) |
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P 6 padding (silent deletion from padded reference) |
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= 7 sequence match |
29
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X 8 sequence mismatch |
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31
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* H can only be present as the first and/or last operation. |
32
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* S may only have H operations between them and the ends of the CIGAR string. |
33
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* For mRNA-to-genome alignment, an N operation represents an intron. For other types of |
34
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alignments, the interpretation of N is not defined. |
35
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* Sum of lengths of the M/I/S/=/X operations shall equal the length of SEQ. |
36
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37
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=head1 EXAMPLES |
38
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39
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# Get the location information on the reference sequence |
40
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$obj_with_role->deletion_positions_on_query; # (10, 19, ...) |
41
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$obj_with_role->insertion_count; # 3 |
42
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$obj_with_role->deletion_positions_on_reference; # (43534511, 43534522, ...) |
43
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44
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=cut |
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46
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# Let the code begin... |
47
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48
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package GenOO::Data::File::SAM::Cigar; |
49
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$GenOO::Data::File::SAM::Cigar::VERSION = '1.5.1'; |
50
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51
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####################################################################### |
52
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####################### Load External modules ##################### |
53
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####################################################################### |
54
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2
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2
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1179
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use Moose::Role; |
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2
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4
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2
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11
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55
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2
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2
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6642
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use namespace::autoclean; |
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3
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2
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13
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56
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57
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58
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####################################################################### |
59
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######################## Required attributes ###################### |
60
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####################################################################### |
61
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requires qw(cigar strand start); |
62
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63
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64
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####################################################################### |
65
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######################## Interface Methods ######################## |
66
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####################################################################### |
67
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sub M_count { |
68
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14
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14
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0
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858
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my ($self) = @_; |
69
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70
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14
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29
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return $self->_total_operator_count('M'); |
71
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} |
72
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73
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sub I_count { |
74
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7
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7
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0
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853
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my ($self) = @_; |
75
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76
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7
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19
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return $self->_total_operator_count('I'); |
77
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} |
78
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79
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sub D_count { |
80
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20
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20
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0
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896
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my ($self) = @_; |
81
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82
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20
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41
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return $self->_total_operator_count('D'); |
83
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} |
84
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85
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sub N_count { |
86
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14
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14
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0
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944
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my ($self) = @_; |
87
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88
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14
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27
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return $self->_total_operator_count('N'); |
89
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} |
90
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91
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sub S_count { |
92
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9
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9
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0
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1095
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my ($self) = @_; |
93
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94
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9
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33
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return $self->_total_operator_count('S'); |
95
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} |
96
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97
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sub H_count { |
98
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1
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1
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0
|
883
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my ($self) = @_; |
99
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100
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1
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7
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return $self->_total_operator_count('H'); |
101
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} |
102
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103
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sub P_count { |
104
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13
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13
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0
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16
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my ($self) = @_; |
105
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106
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13
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23
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return $self->_total_operator_count('P'); |
107
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} |
108
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109
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sub EQ_count { |
110
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14
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14
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0
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925
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my ($self) = @_; |
111
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112
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14
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25
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return $self->_total_operator_count('='); |
113
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} |
114
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115
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sub X_count { |
116
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14
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14
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0
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1088
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my ($self) = @_; |
117
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118
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14
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22
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return $self->_total_operator_count('X'); |
119
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} |
120
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121
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sub insertion_count { |
122
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6
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6
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0
|
1273
|
my ($self) = @_; |
123
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124
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6
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20
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return $self->I_count; |
125
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} |
126
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127
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sub deletion_count { |
128
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6
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6
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0
|
1154
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my ($self) = @_; |
129
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|
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130
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6
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15
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return $self->D_count; |
131
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} |
132
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133
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sub deletion_positions_on_query { |
134
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6
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6
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0
|
1270
|
my ($self) = @_; |
135
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#Tag: AGTGATGGGA------GGATGTCTCGTCTGTGAGTTACAGCA -> CIGAR: 2M1I7M6D26M |
136
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# - - |
137
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#Genome: AG-GCTGGTAGCTCAGGGATGTCTCGTCTGTGAGTTACAGCA -> MD:Z: 3C3T1^GCTCAG26 |
138
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139
|
6
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9
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my @deletion_positions; |
140
|
6
|
100
|
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20
|
if ($self->cigar =~ /D/) { |
141
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5
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16
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my $cigar = $self->cigar_relative_to_query; |
142
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143
|
5
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9
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my $relative_position = 0; |
144
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5
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30
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while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
145
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44
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65
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my $count = $1; |
146
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44
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49
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my $identifier = $2; |
147
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148
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44
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100
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66
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239
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if ($identifier eq 'D') { |
|
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100
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66
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149
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7
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32
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push @deletion_positions, $relative_position - 1; |
150
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} |
151
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elsif ($identifier ne 'N' and $identifier ne 'P' and $identifier ne 'H') { |
152
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35
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138
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$relative_position += $count; |
153
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} |
154
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} |
155
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} |
156
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157
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6
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57
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return @deletion_positions; |
158
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} |
159
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160
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sub mid_position { |
161
|
8
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8
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0
|
782
|
my ($self) = @_; |
162
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# Read: AGTGAT____GGA---GTGACTCA-C -> CIGAR: 2M1I3M4N3M3D1M1I3M1I2M1D1M / 2=1I1=1X1=4N1=1X1=3D1=1I2=1X1I2=1D1= |
163
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# - - - |
164
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# Genome: AG-GCTNNNNGTAGAGG-GAG-CAGC -> MD:Z: 3C1^NNNN1T1^GAG3G2^G1 |
165
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166
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8
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339
|
my $mid_position = $self->start - 1; |
167
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8
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|
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27
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my $mid_position_on_query = ($self->query_length - $self->S_count + 1) / 2; |
168
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8
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24
|
my $cigar = $self->cigar; |
169
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170
|
8
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|
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42
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while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
171
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31
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|
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|
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64
|
my ($count, $identifier) = ($1, $2); |
172
|
31
|
100
|
100
|
|
|
282
|
if ($identifier eq 'D' or $identifier eq 'N' or $identifier eq 'P' or $identifier eq 'H') { |
|
|
100
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66
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50
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66
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100
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100
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173
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6
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26
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$mid_position += $count; |
174
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} |
175
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elsif ($identifier eq 'M' or $identifier eq '=' or $identifier eq 'X') { |
176
|
21
|
100
|
|
|
|
50
|
if ($mid_position_on_query < $count) { |
177
|
7
|
|
|
|
|
51
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return $mid_position + $mid_position_on_query; |
178
|
|
|
|
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} |
179
|
14
|
|
|
|
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16
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$mid_position += $count; |
180
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14
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|
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|
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58
|
$mid_position_on_query -= $count; |
181
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|
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|
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} |
182
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|
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elsif ($identifier eq 'I') { |
183
|
4
|
50
|
|
|
|
12
|
if ($mid_position_on_query < $count) { |
184
|
0
|
|
|
|
|
0
|
return $mid_position + 0.5; |
185
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|
|
|
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} |
186
|
4
|
|
|
|
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19
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$mid_position_on_query -= $count; |
187
|
|
|
|
|
|
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} |
188
|
|
|
|
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} |
189
|
|
|
|
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|
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190
|
1
|
|
|
|
|
8
|
return; |
191
|
|
|
|
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} |
192
|
|
|
|
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|
|
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193
|
|
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194
|
|
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|
|
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sub insertion_positions_on_query { |
195
|
6
|
|
|
6
|
0
|
1253
|
my ($self) = @_; |
196
|
|
|
|
|
|
|
#Tag: AGTGATGGGA------GGATGTCTCGTCTGTGAGTTACAGCA -> CIGAR: 2M1I7M6D26M |
197
|
|
|
|
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# - - |
198
|
|
|
|
|
|
|
#Genome: AG-GCTGGTAGCTCAGGGATGTCTCGTCTGTGAGTTACAGCA -> MD:Z: 3C3T1^GCTCAG26 |
199
|
|
|
|
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|
|
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200
|
6
|
|
|
|
|
8
|
my @insertion_positions; |
201
|
6
|
100
|
|
|
|
21
|
if ($self->cigar =~ /I/) { |
202
|
4
|
|
|
|
|
15
|
my $cigar = $self->cigar_relative_to_query; |
203
|
|
|
|
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|
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204
|
4
|
|
|
|
|
10
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my $relative_position = 0; |
205
|
4
|
|
|
|
|
27
|
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
206
|
41
|
|
|
|
|
63
|
my $count = $1; |
207
|
41
|
|
|
|
|
45
|
my $identifier = $2; |
208
|
|
|
|
|
|
|
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209
|
41
|
100
|
100
|
|
|
268
|
if ($identifier eq 'I') { |
|
|
100
|
66
|
|
|
|
|
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|
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66
|
|
|
|
|
210
|
8
|
|
|
|
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23
|
for (my $i=0; $i<$count; $i++) { |
211
|
8
|
|
|
|
|
11
|
push @insertion_positions, $relative_position; |
212
|
8
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|
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36
|
$relative_position++; |
213
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} |
214
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} |
215
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elsif ($identifier ne 'D' and $identifier ne 'N' and $identifier ne 'P' and $identifier ne 'H') { |
216
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25
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98
|
$relative_position += $count; |
217
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} |
218
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} |
219
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} |
220
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221
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6
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45
|
return @insertion_positions; |
222
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} |
223
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224
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sub mismatch_positions_on_query { |
225
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7
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7
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0
|
795
|
my ($self) = @_; |
226
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#Tag: AGTGATGGGA------GGATGTCTCGTCTGTGAGTTACAGCA -> CIGAR: 2M1I7M6D26M |
227
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# - - |
228
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#Genome: AG-GCTGGTAGCTCAGGGATGTCTCGTCTGTGAGTTACAGCA -> MD:Z: 3C3T1^GCTCAG26 |
229
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230
|
7
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100
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21
|
if ($self->cigar =~ /X/) { |
231
|
1
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2
|
my @mismatch_positions; |
232
|
1
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6
|
my $cigar = $self->cigar_relative_to_query; |
233
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234
|
1
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3
|
my $relative_position = 0; |
235
|
1
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8
|
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
236
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18
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29
|
my $count = $1; |
237
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18
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21
|
my $identifier = $2; |
238
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239
|
18
|
100
|
100
|
|
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139
|
if ($identifier eq 'X') { |
|
|
100
|
66
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66
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240
|
3
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11
|
for (my $i=0; $i<$count; $i++) { |
241
|
3
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5
|
push @mismatch_positions, $relative_position; |
242
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3
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16
|
$relative_position++; |
243
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} |
244
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|
} |
245
|
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|
|
elsif ($identifier ne 'D' and $identifier ne 'N' and $identifier ne 'P' and $identifier ne 'H') { |
246
|
12
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|
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50
|
$relative_position += $count; |
247
|
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|
|
} |
248
|
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|
|
} |
249
|
1
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|
12
|
return @mismatch_positions; |
250
|
|
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|
|
} |
251
|
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|
|
else { |
252
|
6
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22
|
return $self->mismatch_positions_on_query_calculated_from_mdz(); |
253
|
|
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|
|
} |
254
|
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|
|
} |
255
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256
|
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|
|
sub deletion_positions_on_reference { |
257
|
6
|
|
|
6
|
0
|
1241
|
my ($self) = @_; |
258
|
|
|
|
|
|
|
#Tag: AGTGATGGGA------GGATGTCTCGTCTGTGAGTTACAGCA -> CIGAR: 2M1I7M6D26M |
259
|
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|
|
# - - |
260
|
|
|
|
|
|
|
#Genome: AG-GCTGGTAGCTCAGGGATGTCTCGTCTGTGAGTTACAGCA -> MD:Z: 3C3T1^GCTCAG26 |
261
|
|
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|
|
262
|
6
|
|
|
|
|
12
|
my @deletion_positions; |
263
|
6
|
100
|
|
|
|
21
|
if ($self->cigar =~ /D/) { |
264
|
5
|
|
|
|
|
14
|
my $cigar = $self->cigar; |
265
|
|
|
|
|
|
|
|
266
|
5
|
|
|
|
|
8
|
my $relative_position = 0; |
267
|
5
|
|
|
|
|
34
|
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
268
|
44
|
|
|
|
|
73
|
my $count = $1; |
269
|
44
|
|
|
|
|
49
|
my $identifier = $2; |
270
|
|
|
|
|
|
|
|
271
|
44
|
100
|
66
|
|
|
465
|
if ($identifier eq 'D') { |
|
|
100
|
66
|
|
|
|
|
|
|
|
33
|
|
|
|
|
272
|
7
|
|
|
|
|
20
|
for (my $i=0; $i<$count; $i++) { |
273
|
13
|
|
|
|
|
558
|
push @deletion_positions, $self->start + $relative_position; |
274
|
13
|
|
|
|
|
63
|
$relative_position++; |
275
|
|
|
|
|
|
|
} |
276
|
|
|
|
|
|
|
} |
277
|
|
|
|
|
|
|
elsif ($identifier ne 'I' and $identifier ne 'P' and $identifier ne 'S' and $identifier ne 'H') { |
278
|
29
|
|
|
|
|
123
|
$relative_position += $count; |
279
|
|
|
|
|
|
|
} |
280
|
|
|
|
|
|
|
} |
281
|
|
|
|
|
|
|
} |
282
|
|
|
|
|
|
|
|
283
|
6
|
|
|
|
|
53
|
return @deletion_positions; |
284
|
|
|
|
|
|
|
} |
285
|
|
|
|
|
|
|
|
286
|
|
|
|
|
|
|
sub mismatch_positions_on_reference { |
287
|
20
|
|
|
20
|
0
|
1254
|
my ($self) = @_; |
288
|
|
|
|
|
|
|
#Tag: AGTGATGGGA------GGATGTCTCGTCTGTGAGTTACAGCA -> CIGAR: 2M1I7M6D26M |
289
|
|
|
|
|
|
|
# - - |
290
|
|
|
|
|
|
|
#Genome: AG-GCTGGTAGCTCAGGGATGTCTCGTCTGTGAGTTACAGCA -> MD:Z: 3C3T1^GCTCAG26 |
291
|
|
|
|
|
|
|
|
292
|
20
|
100
|
|
|
|
48
|
if ($self->cigar =~ /X/) { |
293
|
2
|
|
|
|
|
4
|
my @mismatch_positions; |
294
|
2
|
|
|
|
|
7
|
my $cigar = $self->cigar; |
295
|
|
|
|
|
|
|
|
296
|
2
|
|
|
|
|
5
|
my $relative_position = 0; |
297
|
2
|
|
|
|
|
16
|
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
298
|
36
|
|
|
|
|
47
|
my $count = $1; |
299
|
36
|
|
|
|
|
31
|
my $identifier = $2; |
300
|
|
|
|
|
|
|
|
301
|
36
|
100
|
66
|
|
|
206
|
if ($identifier eq 'X') { |
|
|
100
|
66
|
|
|
|
|
|
|
|
33
|
|
|
|
|
302
|
6
|
|
|
|
|
15
|
for (my $i=0; $i<$count; $i++) { |
303
|
6
|
|
|
|
|
190
|
push @mismatch_positions, $self->start + $relative_position; |
304
|
6
|
|
|
|
|
27
|
$relative_position++; |
305
|
|
|
|
|
|
|
} |
306
|
|
|
|
|
|
|
} |
307
|
|
|
|
|
|
|
elsif ($identifier ne 'I' and $identifier ne 'P' and $identifier ne 'S' and $identifier ne 'H') { |
308
|
24
|
|
|
|
|
76
|
$relative_position += $count; |
309
|
|
|
|
|
|
|
} |
310
|
|
|
|
|
|
|
} |
311
|
2
|
|
|
|
|
15
|
return @mismatch_positions; |
312
|
|
|
|
|
|
|
} |
313
|
|
|
|
|
|
|
else { |
314
|
18
|
|
|
|
|
55
|
return $self->mismatch_positions_on_reference_calculated_from_mdz(); |
315
|
|
|
|
|
|
|
} |
316
|
|
|
|
|
|
|
} |
317
|
|
|
|
|
|
|
|
318
|
|
|
|
|
|
|
sub cigar_relative_to_query { |
319
|
15
|
|
|
15
|
0
|
796
|
my ($self) = @_; |
320
|
|
|
|
|
|
|
|
321
|
15
|
|
|
|
|
41
|
my $cigar = $self->cigar; |
322
|
|
|
|
|
|
|
|
323
|
|
|
|
|
|
|
# if negative strand -> reverse the cigar string |
324
|
15
|
100
|
100
|
|
|
524
|
if (defined $self->strand and ($self->strand == -1)) { |
325
|
4
|
|
|
|
|
8
|
my $reverse_cigar = ''; |
326
|
4
|
|
|
|
|
23
|
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
327
|
16
|
|
|
|
|
64
|
$reverse_cigar = $1.$2.$reverse_cigar; |
328
|
|
|
|
|
|
|
} |
329
|
4
|
|
|
|
|
17
|
return $reverse_cigar; |
330
|
|
|
|
|
|
|
} |
331
|
|
|
|
|
|
|
else { |
332
|
11
|
|
|
|
|
36
|
return $cigar; |
333
|
|
|
|
|
|
|
} |
334
|
|
|
|
|
|
|
} |
335
|
|
|
|
|
|
|
|
336
|
|
|
|
|
|
|
sub aligned_areas_starts_and_stops { |
337
|
0
|
|
|
0
|
0
|
0
|
my ($self) = @_; |
338
|
|
|
|
|
|
|
|
339
|
|
|
|
|
|
|
# returns the start and stop positions on the reference |
340
|
|
|
|
|
|
|
# for all parts of the alignment that bind. |
341
|
|
|
|
|
|
|
|
342
|
|
|
|
|
|
|
# Read: AGTGAT____GGA---GTGACTCA-C -> CIGAR: 2M1I3M4N3M3D1M1I3M1I2M1D1M / 2=1I1=1X1=4N1=1X1=3D1=1I2=1X1I2=1D1= |
343
|
|
|
|
|
|
|
# - - - |
344
|
|
|
|
|
|
|
# Genome: AG-GCTNNNNGTAGAGG-GAG-CAGC -> MD:Z: 3C1^NNNN1T1^GAG3G2^G1 |
345
|
|
|
|
|
|
|
|
346
|
0
|
|
|
|
|
0
|
my @exon_starts; |
347
|
|
|
|
|
|
|
my @exon_stops; |
348
|
|
|
|
|
|
|
|
349
|
0
|
|
|
|
|
0
|
my $pos = $self->start; |
350
|
0
|
|
|
|
|
0
|
my $cigar = $self->cigar; |
351
|
|
|
|
|
|
|
|
352
|
0
|
|
|
|
|
0
|
while ($cigar =~ /(\d+)([MIDNSHP=X])/g) { |
353
|
0
|
|
|
|
|
0
|
my ($count, $identifier) = ($1, $2); |
354
|
0
|
0
|
0
|
|
|
0
|
if ($identifier eq 'D' or $identifier eq 'N' or $identifier eq 'P' or $identifier eq 'H') { |
|
|
0
|
0
|
|
|
|
|
|
|
|
0
|
|
|
|
|
|
|
|
0
|
|
|
|
|
|
|
|
0
|
|
|
|
|
355
|
0
|
|
|
|
|
0
|
$pos += $count; |
356
|
|
|
|
|
|
|
} |
357
|
|
|
|
|
|
|
elsif ($identifier eq 'M' or $identifier eq '=' or $identifier eq 'X') { |
358
|
0
|
|
|
|
|
0
|
push @exon_starts, $pos; |
359
|
0
|
|
|
|
|
0
|
push @exon_stops, $pos+$count-1; |
360
|
0
|
|
|
|
|
0
|
$pos += $count; |
361
|
|
|
|
|
|
|
} |
362
|
|
|
|
|
|
|
} |
363
|
0
|
|
|
|
|
0
|
return (\@exon_starts, \@exon_stops); |
364
|
|
|
|
|
|
|
} |
365
|
|
|
|
|
|
|
|
366
|
|
|
|
|
|
|
####################################################################### |
367
|
|
|
|
|
|
|
######################### Private methods ########################## |
368
|
|
|
|
|
|
|
####################################################################### |
369
|
|
|
|
|
|
|
sub _total_operator_count { |
370
|
106
|
|
|
106
|
|
122
|
my ($self, $operator) = @_; |
371
|
|
|
|
|
|
|
|
372
|
106
|
|
|
|
|
221
|
my $cigar = $self->cigar; |
373
|
|
|
|
|
|
|
|
374
|
106
|
|
|
|
|
97
|
my $count = 0; |
375
|
106
|
|
|
|
|
996
|
while ($cigar =~ /(\d+)$operator/g) { |
376
|
115
|
|
|
|
|
446
|
$count += $1; |
377
|
|
|
|
|
|
|
} |
378
|
106
|
|
|
|
|
637
|
return $count; |
379
|
|
|
|
|
|
|
} |
380
|
|
|
|
|
|
|
|
381
|
|
|
|
|
|
|
1; |