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 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238
|
#!/usr/bin/env perl
use strict;
use warnings;
use lib ($ENV{EUK_MODULES});
use Fasta_reader;
use SAM_reader;
use SAM_entry;
use Data::Dumper;
my $FASTA_LENGTH = 60;
my $usage = "usage: $0 alignments.sam target.fasta [JUST_ALIGN_STATS=0]\n\n";
my $sam_alignments = $ARGV[0] or die $usage;
my $target_fasta = $ARGV[1] or die $usage;
my $JUST_STATS = $ARGV[2] || 0;
main: {
my $fasta_reader = new Fasta_reader($target_fasta);
my %fasta_seqs = $fasta_reader->retrieve_all_seqs_hash();
my $sam_reader = new SAM_reader($sam_alignments);
my $curr_scaff_acc = "";
my $curr_scaff_seq = "";
while (my $sam_entry = $sam_reader->get_next() ) {
my $read_name = $sam_entry->reconstruct_full_read_name();
my $read_seq = uc $sam_entry->get_sequence();
my $scaff_name = $sam_entry->get_scaffold_name();
if ($scaff_name eq "*") { next; } # not aligned.
if ($scaff_name ne $curr_scaff_acc) {
$curr_scaff_acc = $scaff_name;
$curr_scaff_seq = uc $fasta_seqs{$scaff_name};
}
my ($genome_coords_aref, $read_coords_aref) = $sam_entry->get_alignment_coords();
unless (scalar(@$genome_coords_aref) >= 1) { next; }
unless ($JUST_STATS) {
## coordinate dump.
print "// $scaff_name (top) vs. $read_name (bottom):\n\n";
print "$scaff_name length: " . length($curr_scaff_seq) . "\n";
print "$read_name length: " . length($read_seq) . "\n";
print "$scaff_name\t$read_name\n";
for (my $i = 0; $i <= $#$genome_coords_aref; $i++) {
my ($genome_lend, $genome_rend) = @{$genome_coords_aref->[$i]};
my ($read_lend, $read_rend) = @{$read_coords_aref->[$i]};
print "$genome_lend-$genome_rend\t$read_lend-$read_rend\n";
}
print "\n";
}
eval {
&draw_alignment($scaff_name, $read_name, $genome_coords_aref, $read_coords_aref, \$curr_scaff_seq, \$read_seq);
};
if ($@) {
print STDERR "$@\n";
}
}
exit(0);
}
####
sub draw_alignment {
my ($scaff_name, $read_name, $genome_coords_aref, $read_coords_aref, $curr_scaff_seq_sref, $read_seq_sref) = @_;
my $scaff_align_string = "";
my $read_align_string = "";
my $prev_scaff_coord = 0;
my $prev_read_coord = 0;
my $num_indels = scalar(@$genome_coords_aref) - 1;
my $num_aligned_bases = 0;
while (@$genome_coords_aref) {
my $genome_coordset = shift @$genome_coords_aref;
my $read_coordset = shift @$read_coords_aref;
my ($genome_lend, $genome_rend) = @$genome_coordset;
my ($read_lend, $read_rend) = @$read_coordset;
## sometimes bwasw is off at the end of the contig
if ($genome_rend > length($$curr_scaff_seq_sref)) {
my $delta = $genome_rend - length($$curr_scaff_seq_sref);
$genome_rend -= $delta;
$read_rend -= $delta;
# print STDERR "*adjusting by $delta\n";
}
$num_aligned_bases += $read_rend - $read_lend + 1;
if ($prev_read_coord > 0 && $read_lend - $prev_read_coord > 1) {
## add gaps to read string
for (my $i = $prev_read_coord + 1; $i < $read_lend; $i++) {
$read_align_string .= substr($$read_seq_sref, $i-1, 1);
$scaff_align_string .= "-";;
}
}
if ($prev_scaff_coord > 0 && $genome_lend - $prev_scaff_coord > 1) {
# add gaps to genome string
for (my $i = $prev_scaff_coord + 1; $i < $genome_lend; $i++) {
$scaff_align_string .= substr($$curr_scaff_seq_sref, $i-1, 1);
$read_align_string .= "-";
}
}
$prev_read_coord = $read_rend;
$prev_scaff_coord = $genome_rend;
my $read_seq_region_len = $read_rend - $read_lend + 1;
my $genome_seq_region_len = $genome_rend - $genome_lend + 1;
my $read_seq_region = substr($$read_seq_sref, $read_lend - 1, $read_rend - $read_lend + 1);
my $genome_seq_region = substr($$curr_scaff_seq_sref, $genome_lend - 1, $genome_rend - $genome_lend + 1);
if (length($read_seq_region) != length($genome_seq_region)) {
die "Error, lengths of regions are different: ($read_seq_region_len vs. $genome_seq_region_len) extracted:\n"
. "read_seq_region:\t" . $read_seq_region . "\n"
. "genome_seq_regn:\t" . $genome_seq_region . "\n";
}
$scaff_align_string .= $genome_seq_region;
$read_align_string .= $read_seq_region;
}
my $alignment_length = length($scaff_align_string);
my $num_gaps = 0;
while ($scaff_align_string =~ /-/g) { $num_gaps++; }
while ($read_align_string =~ /-/g) { $num_gaps++; }
my $percent_gap = $num_gaps / $alignment_length * 100;
eval {
my ($num_mismatches) = &print_pretty($scaff_align_string, $read_align_string);
print join("\t", "#", "scaff_name", "read_name", "read_length", "aligned_bases", "matches", "mismatches", "indel_bkpts", "sum_indel_lens", "pct_mismatches", "pct_indel_bkpts", "pct_indel_lens") . "\n";
print join("\t", "#", $scaff_name, $read_name, length($$read_seq_sref),
$num_aligned_bases, $num_aligned_bases - $num_mismatches, $num_mismatches, $num_indels, $num_gaps,
sprintf("%.2f", $num_mismatches/$num_aligned_bases*100),
sprintf("%.2f", $num_indels/$num_aligned_bases*100),
sprintf("%.2f", $percent_gap),
) . "\n\n\n";
};
if ($@) {
print STDERR "$@\n";
die;
}
return;
}
####
sub print_pretty {
my ($scaff_align_string, $read_align_string) = @_;
if (length($scaff_align_string) != length($read_align_string) ) {
die "Error, alignment lengths differ.:\n"
. "Scaff_align_string: " . length($scaff_align_string) . "\t$scaff_align_string\n"
. "Read_align_string: " . length($read_align_string) . "\t$read_align_string\n";
}
my @scaff_align_chars = split(//, $scaff_align_string);
my @read_align_chars = split(//, $read_align_string);
my $top_text = "";
my $match_text = "";
my $bottom_text = "";
my $num_mismatches = 0;
for (my $i = 0; $i <= $#scaff_align_chars; $i++) {
if ($i != 0 && $i % 60 == 0) {
print join("\n", $top_text, $match_text, $bottom_text) . "\n\n" unless $JUST_STATS;
$top_text = ""; $match_text = ""; $bottom_text = "";
}
my $match_char = ".";
if ($scaff_align_chars[$i] ne "-" && $read_align_chars[$i] ne "-") {
$match_char = ($scaff_align_chars[$i] eq $read_align_chars[$i]) ? "|" : "*";
if ($match_char eq "*") {
$num_mismatches++;
}
}
$top_text .= $scaff_align_chars[$i];
$bottom_text .= $read_align_chars[$i];
$match_text .= $match_char;
}
if ($top_text) {
print join("\n", $top_text, $match_text, $bottom_text) . "\n\n" unless $JUST_STATS;
}
return ($num_mismatches);
}
|