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+line | stmt | bran | cond | sub | time | code |
+1 | | | | | | #!/usr/bin/env perl |
+2 | | | | | | return 1 if caller(); #tests |
+3 | |
4 | 3 3 3 3 3 3 3 3 3 3 3 3 | | | | 2670 3 93 8 4 55 8 2 13 49 3 128 | use strict; use warnings; use diagnostics; use feature qw(say); |
+5 | 3 3 3 3 3 3 | | | | 3975 19485 13 2237 145624 534 | use Getopt::Long; use Pod::Usage; |
+6 | 3 3 3 | | | | 28 9 120 | use Readonly; |
+7 | 3 3 3 3 3 3 | | | | 10 1 65 50 3 31 | use Bio::Seq; use Bio::SeqIO; |
+8 | 3 3 3 3 3 3 | | | | 43 2 120 12 2 23 | use FindBin; use lib "$FindBin::RealBin/lib"; |
+9 | 3 3 3 3 3 3 | | | | 328 3 70 11 1 68 | use Search; use SS; # lib modules |
+10 | | | | | | # Own Modules (https://github.com/bretonics/Modules) |
+11 | 3 3 3 3 3 3 3 3 3 | | | | 5 4 121 10 3 92 8 5 4615 | use MyConfig; use MyIO; use Handlers; |
+12 | |
13 | | | | | | # ============================================================================== |
+14 | | | | | | # |
+15 | | | | | | # CAPITAN: Andres Breton, http://andresbreton.com |
+16 | | | | | | # FILE: chomp.pl |
+17 | | | | | | # LICENSE: MIT |
+18 | | | | | | # USAGE: Find guide RNA (gRNA) sequences for CRISPR targeting |
+19 | | | | | | # DEPENDENCIES: - BioPerl modules |
+20 | | | | | | # - Own 'Modules' repo |
+21 | | | | | | # |
+22 | | | | | | # ============================================================================== |
+23 | |
24 | |
25 | | | | | | #------------------------------------------------------------------------------- |
+26 | | | | | | # COMMAND LINE |
+27 | | | | | | my $SEQ; |
+28 | | | | | | my @SUBJECTS; |
+29 | | | | | | my $DOWNSEQ; |
+30 | | | | | | my $UPSEQ; |
+31 | | | | | | my $SS; |
+32 | | | | | | my $OUTFILE; |
+33 | | | | | | my $VERBOSE; |
+34 | |
35 | | | | | | our $OUTDIR = 'gRNAs'; |
+36 | | | | | | our $WINDOWSIZE = 23; |
+37 | |
38 | |
39 | | | | | | my $USAGE = "\n\n$0 [options]\n |
+40 | | | | | | Options: |
+41 | | | | | | -seq Sequence file to search gRNAs [required] |
+42 | | | | | | -subjects Subject sequence file(s) to BLAST search (search instead of -seq) |
+43 | | | | | | -down Down sequence to append |
+44 | | | | | | -up Up sequence to append |
+45 | | | | | | -window Window size for gRNA sequence (default = 23) |
+46 | | | | | | -ss Secondary structure prediction |
+47 | | | | | | -out Out file name [required] |
+48 | | | | | | -outdir Out directory name |
+49 | | | | | | -help Shows this message |
+50 | | | | | | \n"; |
+51 | |
52 | | | | | | # OPTIONS |
+53 | | | | | | GetOptions( |
+54 | | | | | | 'seq=s' =>\$SEQ, |
+55 | | | | | | 'subjects:s{,10}' =>\@SUBJECTS, |
+56 | | | | | | 'down:s' =>\$DOWNSEQ, |
+57 | | | | | | 'up:s' =>\$UPSEQ, |
+58 | | | | | | 'window:i' =>\$WINDOWSIZE, |
+59 | | | | | | 'ss!' =>\$SS, |
+60 | | | | | | 'out=s' =>\$OUTFILE, |
+61 | | | | | | 'outdir:s' =>\$OUTDIR, |
+62 | | | | | | 'verbose!' =>\$VERBOSE, |
+63 | | | | | | help =>sub{pod2usage($USAGE);} |
+64 | | | | | | )or pod2usage(2); |
+65 | |
66 | | | | | | checks(); # check CL arguments |
+67 | |
68 | | | | | | #------------------------------------------------------------------------------- |
+69 | | | | | | # VARIABLES |
+70 | | | | | | my $AUTHOR = 'Andres Breton, <dev@andresbreton.com>'; |
+71 | |
72 | | | | | | my $REALBIN = $FindBin::RealBin; |
+73 | |
74 | | | | | | my ($seqDetails) = getSeqDetails($SEQ); |
+75 | | | | | | my @SUBJSEQS; # sequence file to use in BLAST search |
+76 | |
77 | | | | | | #------------------------------------------------------------------------------- |
+78 | | | | | | # CALLS |
+79 | | | | | | mkDir($OUTDIR); |
+80 | | | | | | my $gRNAs = findOligo($seqDetails, $WINDOWSIZE); # gRNA HoH |
+81 | | | | | | my $fastaFile = writeFasta($gRNAs, $OUTFILE); # Write gRNAs fasta file |
+82 | | | | | | my $targets = Search::blast($fastaFile, \@SUBJSEQS, $OUTFILE); # gRNA target hits |
+83 | | | | | | writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE); |
+84 | |
85 | | | | | | #------------------------------------------------------------------------------- |
+86 | | | | | | # SUBS |
+87 | |
88 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+89 | | | | | | # $input = checks(); |
+90 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+91 | | | | | | # This function checks for arguments passed on the command-line using global variables. |
+92 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+93 | | | | | | # $return = Prompts users and exits if errors |
+94 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+95 | | | | | | sub checks { |
+96 | | | | | | # Command line arguments passed |
+97 | |
98 | | | | | | # Exits |
+99 | 3 | | | | 212 | die 'Did not provide an input file, -seq <infile>', $USAGE unless ($SEQ); |
+100 | 0 | | | | 0 | die 'Did not provide an output file, -out <outfile>', $USAGE unless ($OUTFILE) ; |
+101 | |
102 | | | | | | # Warnings |
+103 | 0 | | | | 0 | say 'Did not provide a DOWN stream sequence to append to gRNA seq, -down <seq>' unless ($DOWNSEQ && $VERBOSE); |
+104 | 0 | | | | 0 | say 'Did not provide an UP stream sequence to append to gRNA seq, -up <seq>' unless ($UPSEQ && $VERBOSE); |
+105 | |
106 | |
107 | 0 | | | | 0 | setParameters(); # set parameters to use in calls |
+108 | |
109 | 0 | | | | 0 | return; |
+110 | | | | | | } |
+111 | |
112 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+113 | | | | | | # $input = (); |
+114 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+115 | | | | | | # This function takes no arguments, uses global variables to decide which set of |
+116 | | | | | | # variables to use and set for calls. |
+117 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+118 | | | | | | # $output = sets sequence file to use for BLAST search |
+119 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+120 | | | | | | sub setParameters { |
+121 | | | | | | # Set up Variables |
+122 | 3 | | | | 13 | if (@SUBJECTS) { |
+123 | 0 | | | | 0 | @SUBJSEQS = @SUBJECTS; |
+124 | | | | | | } elsif ($SEQ) { |
+125 | 0 | | | | 0 | @SUBJSEQS = $SEQ; |
+126 | | | | | | } else { |
+127 | 3 | | | | 21 | die 'Could not determine which file(s) to use as search sequence.' |
+128 | | | | | | } |
+129 | |
130 | 0 0 | | | | 0 0 | mkDir($OUTDIR); mkDir("$OUTDIR/ss") if($SS); |
+131 | |
132 | 0 | | | | 0 | return; |
+133 | | | | | | } |
+134 | |
135 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+136 | | | | | | # $input = (); |
+137 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+138 | | | | | | # This function takes 5 arguments; gRNAs HoH, BLAST target HoA containing matches |
+139 | | | | | | # throughout the sequence, the down/up stream sequences to append to gRNA |
+140 | | | | | | # sequences, and the output file name |
+141 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+142 | | | | | | # $output = File containing gRNAs sequences and relative information on results |
+143 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+144 | | | | | | sub writeResults { |
+145 | 36 | | | | 101 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '(\%gRNAs, \%targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE)'; |
+146 | 36 | | | | 1531 | @_ == 6 or die wrongNumberArguments(), $filledUsage; |
+147 | |
148 | 18 | | | | 54 | my ($gRNAs, $targets, $down, $up, $window, $file) = @_; |
+149 | |
150 | 18 | | | | 153 | my %targets = %$targets; |
+151 | 15 | | | | 25 | my $num = keys %$gRNAs; # number of gRNA sequences |
+152 | 15 | | | | 35 | my $outFile = "$OUTDIR/$file.txt"; |
+153 | |
154 | 15 | | | | 67 | my $FH = getFH(">", $outFile); |
+155 | 15 | | | | 2556 | say $FH "Name\tSequence\tStrand\tPalindrome\tSubject\tStart\tOccurrences\tIdentities"; |
+156 | |
157 | 15 | | | | 50 | my ($subjects, $sortedgRNAs, $details) = sortResults(\%targets); |
+158 | 15 | | | | 18 | my @subjects = @$subjects; |
+159 | 15 | | | | 27 | my %sortedgRNAs = %$sortedgRNAs; |
+160 | 15 | | | | 49 | my %details = %$details; |
+161 | |
162 | |
163 | | | | | | # Get ordered gRNA sequences + info to print |
+164 | 15 | | | | 25 | foreach my $subject (@subjects) { |
+165 | 10 10 | | | | 14 188 | foreach my $gRNA ( @{$sortedgRNAs{$subject}} ) { |
+166 | 170 | | | | 154 | my $sequence = $gRNAs->{$gRNA}->{'sequence'}; |
+167 | 170 | | | | 115 | my $palindrome = $gRNAs->{$gRNA}{'palindrome'}; |
+168 | |
169 | | | | | | # Complete oligo sequence: |
+170 | | | | | | # + DOWN flanking target region |
+171 | | | | | | # + gRNA sequence |
+172 | | | | | | # + UP flanking target region |
+173 | 170 | | | | 519 | if (!$down and !$up) { |
+174 | 34 | | | | 14 | $sequence = $sequence; |
+175 | | | | | | } elsif ($down and !$up) { |
+176 | 34 | | | | 22 | $sequence = $down . $sequence; |
+177 | | | | | | } elsif (!$down and $up) { |
+178 | 34 | | | | 23 | $sequence = $sequence . $up; |
+179 | | | | | | } else { |
+180 | 68 | | | | 31 | $sequence = $down . $sequence . $up; |
+181 | | | | | | } |
+182 | |
183 | | | | | | # Get all details to print to file |
+184 | 170 | | | | 124 | my $position = $gRNAs->{$gRNA}{'start'}; # gRNA sequence position |
+185 | 170 | | | | 87 | my $strand = $gRNAs->{$gRNA}{'strand'}; |
+186 | 170 | | | | 112 | my $occurrence = $details->{$gRNA}{$subject}->{'occurrences'}; |
+187 | 170 170 | | | | 62 138 | my $identities = join("," , @{ $details->{$gRNA}{$subject}->{'unqIdentities'} } ); # get string of identities |
+188 | 170 170 | | | | 69 185 | my $sStart = @{ $targets{$gRNA}{$subject}{'hsps'} }[0]->{'sstart'}; # get location of BLAST match hit in subject (reference) for gRNA found |
+189 | |
190 | 170 | | | | 209 | say $FH "$gRNA\t$sequence\t$strand\t$palindrome\t$subject\t$sStart\t$occurrence\t$identities"; # print to file |
+191 | | | | | | } |
+192 | 15 | | | | 5663 | } close $FH; |
+193 | |
194 | 15 | | | | 242 | say "gRNAs file written to $outFile"; |
+195 | 15 | | | | 752 | return; |
+196 | | | | | | } |
+197 | | | | | | #------------------------------------------------------------------------------- |
+198 | | | | | | # HELPERS |
+199 | |
200 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+201 | | | | | | # $input = ($seqFile); |
+202 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+203 | | | | | | # This function takes 1 argument, the sequence file to get details using |
+204 | | | | | | # BioPerl modules |
+205 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+206 | | | | | | # $return = ( \%seqInfo ); Returns sequence details hash |
+207 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+208 | | | | | | sub getSeqDetails { |
+209 | 18 | | | | 51 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '($seqFile)'; |
+210 | 18 | | | | 701 | @_ == 1 or die wrongNumberArguments(), $filledUsage; |
+211 | |
212 | 12 | | | | 16 | my ($seqFile) = @_; |
+213 | |
214 | | | | | | # Sequence OO |
+215 | 12 | | | | 95 | my $seqInObj = Bio::SeqIO->new(-file => $seqFile, -alphabet => "dna"); |
+216 | 6 | | | | 3002 | my $format = $seqInObj->_guess_format($seqFile); #check format of input file |
+217 | 6 | | | | 88 | my $seqObj = $seqInObj->next_seq; |
+218 | 6 | | | | 1215 | my $sequence = uc $seqObj->seq; |
+219 | 6 | | | | 102 | my $reverse = uc $seqObj->revcom->seq; |
+220 | |
221 | 6 | | | | 929 | my %seqInfo = ( |
+222 | | | | | | 'seqInObj' => $seqInObj, |
+223 | | | | | | 'format' => $format, |
+224 | | | | | | 'seqObj' => $seqObj, |
+225 | | | | | | 'sequence' => $sequence, |
+226 | | | | | | 'reverse' => $reverse, |
+227 | | | | | | ); |
+228 | |
229 | 6 | | | | 99 | return(\%seqInfo); |
+230 | | | | | | } |
+231 | |
232 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+233 | | | | | | # $input = ($gRNAs, $OUTDIR, $OUTFILE); |
+234 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+235 | | | | | | # This function takes 3 arguments; HoH reference of gRNA sequences, |
+236 | | | | | | # the output diretory, and the output file name. Writes each gRNA |
+237 | | | | | | # sequence found in FASTA and returns file location. |
+238 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+239 | | | | | | # $return = ($outFile); |
+240 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+241 | | | | | | sub writeFasta { |
+242 | 21 | | | | 65 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '($gRNAs, $OUTFILE)'; |
+243 | 21 | | | | 915 | @_ == 2 or die wrongNumberArguments(), $filledUsage; |
+244 | |
245 | 12 | | | | 21 | my ($gRNAs, $out) = @_; |
+246 | 12 | | | | 29 | my $outFile = "$OUTDIR/$out.fasta"; |
+247 | 12 | | | | 53 | my $FH = getFH(">", $outFile); |
+248 | 12 | | | | 2096 | my $num = keys %$gRNAs; # number of gRNA sequences |
+249 | |
250 | 6 | | | | 35 | for (my $i = 0; $i < $num; $i++) { # get sequences in numerical order |
+251 | 102 | | | | 72 | my $name = "gRNA_$i"; |
+252 | 102 | | | | 138 | my $sequence = $gRNAs->{$name}->{'sequence'}; |
+253 | 102 | | | | 87 | ss($name, $sequence) if($SS); # secondary structure prediction if desired |
+254 | 102 | | | | 202 | say $FH ">$name:" . $gRNAs->{$name}->{'start'}; # 'gRNA_0:start_position' |
+255 | 102 | | | | 135 | say $FH $sequence; |
+256 | 6 | | | | 3288 | } close $FH; |
+257 | |
258 | 6 | | | | 153 | return $outFile; |
+259 | | | | | | } |
+260 | |
261 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+262 | | | | | | # $input = ($targetsRef); |
+263 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+264 | | | | | | # This function takes 1 arguments: $target hash of array of hashes |
+265 | | | | | | # references returned from Search::blast. Returns a numerically |
+266 | | | | | | # ordered array of gRNA sequence names based on lowest identity |
+267 | | | | | | # base pair matches, then occurrences and a details hash with sorted |
+268 | | | | | | # identities and number of occurrences per gRNA sequence. |
+269 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+270 | | | | | | # $return = ( \@sortedgRNAs, \%details ); |
+271 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+272 | | | | | | sub sortResults { |
+273 | 15 | | | | 33 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '($targetsRef)'; |
+274 | 15 | | | | 418 | @_ == 1 or die wrongNumberArguments(), $filledUsage; |
+275 | |
276 | 15 | | | | 20 | my ($targetsRef) = @_; |
+277 | | | | | | # %targets HoHoAoH: |
+278 | | | | | | # -- Hash key == gRNA name |
+279 | | | | | | # -- Hash key == Subject name |
+280 | | | | | | # -- Hash key == 'info' |
+281 | | | | | | # -- Hash key == 'hsps' |
+282 | | | | | | # -- Array accounts for multiple hits for each gRNA sequence |
+283 | | | | | | # -- Hash contains BLAST match info |
+284 | 15 | | | | 70 | my %targets = %$targetsRef; |
+285 | 15 | | | | 50 | my @gRNAs = keys %targets; |
+286 | 15 | | | | 21 | my (@subjects, @noHits, @sorted, %sortedgRNAs, %details); |
+287 | |
288 | | | | | | # Get number of occurrences from BLAST call per gRNA sequence in %targets Hash of Hashes of Array of Hashes |
+289 | 15 | | | | 25 | foreach my $gRNA (@gRNAs) { # iterate through each gRNA instance |
+290 | 170 | | | | 237 | @subjects = sort keys $targets{$gRNA}; # @subjects == BLAST subject instances in Hash of Arrays of Hash |
+291 | |
292 | 170 | | | | 106 | foreach my $subject (@subjects) { # iterate through each BLAST subject instance |
+293 | | | | | | # Handle gRNA sequences having 'No hits', skip sorting |
+294 | 170 170 | | | | 86 301 | next if ( @{ $targets{$gRNA}{$subject}{'info'} }[0]->{'numhits'} == 0 ); |
+295 | |
296 | 170 | | | | 88 | my @identities; |
+297 | 170 | | | | 212 | my @ids = sortIdentities( $targets{$gRNA}{$subject}{'hsps'} ); # get sorted list of all BLAST hits (and for all subjects) for each gRNA query |
+298 | 170 | | | | 196 | push @identities, @ids; # push identities list for each subject |
+299 | |
300 | | | | | | # Remove duplicates |
+301 | 170 | | | | 62 | my @unqIdentities; |
+302 | 170 | | | | 95 | foreach my $value (@identities) { |
+303 | | | | | | # Push identitities value unless already recorded |
+304 | 460 620 | | | | 288 409 | next if ( grep { $_ == $value} @unqIdentities ); |
+305 | 310 | | | | 247 | push @unqIdentities, $value; |
+306 | | | | | | } |
+307 | |
308 | | | | | | # Sort identities from all HSPs |
+309 | 170 480 | | | | 121 193 | @identities = ( sort {$b <=> $a} @identities ); |
+310 | 170 210 | | | | 95 91 | @unqIdentities = ( sort {$b <=> $a} @unqIdentities ); |
+311 | |
312 | | | | | | # Number of hashes in array == number of matches for same gRNA sequence throughout the whole sequence |
+313 | 170 | | | | 91 | my $occurrences = @identities; # number of occcurrences per gRNA |
+314 | |
315 | | | | | | # %details |
+316 | | | | | | # -- Hash key == gRNA name |
+317 | | | | | | # -- Hash key == subject |
+318 | | | | | | # -- Hash keys == 'identities', 'unique identities', and 'number of occurrences' |
+319 | 170 | | | | 469 | $details{$gRNA}{$subject} = { |
+320 | | | | | | 'identities' => \@identities, |
+321 | | | | | | 'unqIdentities' => \@unqIdentities, |
+322 | | | | | | 'occurrences' => $occurrences, |
+323 | | | | | | }; |
+324 | | | | | | } |
+325 | | | | | | } |
+326 | |
327 | | | | | | # Return sorted gRNA names based on lowest identity base pair matches, then occurrences |
+328 | 15 | | | | 21 | foreach my $subject ( @subjects ) { |
+329 | | | | | | # Get only gRNAs having hits in current subject, remove others |
+330 | 10 170 170 | | | | 12 63 160 | my @gRNAsHit = grep { @{ $targets{$_}{$subject}{'info'} }[0]->{'numhits'} != 0 } @gRNAs; |
+331 | |
332 | 10 505 | | | | 11 811 | my @sorted = ( sort { $details{$a}{$subject}{'unqIdentities'}[0] <=> $details{$b}{$subject}{'unqIdentities'}[0] || $details{$a}{$subject}{'occurrences'} <=> $details{$b}{$subject}{'occurrences'} } @gRNAsHit ); |
+333 | 10 | | | | 21 | $sortedgRNAs{$subject} = \@sorted; |
+334 | | | | | | } |
+335 | |
336 | 15 | | | | 46 | return (\@subjects, \%sortedgRNAs, \%details); |
+337 | | | | | | } |
+338 | |
339 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+340 | | | | | | # $input = ($targets{$name}); |
+341 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+342 | | | | | | # This function takes 1 argument: an array reference containing |
+343 | | | | | | # all gRNA matches for a given gRNA sequence name |
+344 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+345 | | | | | | # $return = sorted identities ascending numerically |
+346 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+347 | | | | | | sub sortIdentities { |
+348 | 182 | | | | 195 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '($targets{$gRNA}{$subject}{\'hsps\'})'; |
+349 | 182 | | | | 3737 | @_ == 1 or die wrongNumberArguments(), $filledUsage; |
+350 | |
351 | 176 | | | | 100 | my ($hsps) = @_; |
+352 | 176 | | | | 193 | my @hsps = @$hsps; |
+353 | 172 | | | | 85 | my $numMatches = @hsps; |
+354 | 172 | | | | 79 | my @identities; |
+355 | |
356 | 172 | | | | 105 | foreach my $hash (@hsps) { |
+357 | 620 620 | | | | 437 265 | my $nident = $hash->{'nident'}; chomp($nident); |
+358 | 620 | | | | 445 | return ($nident) if ($numMatches == 1); # return single match (when $nident == $WINDOWSIZE is only match) |
+359 | 600 | | | | 661 | push @identities, $nident unless ($nident == $WINDOWSIZE); |
+360 | | | | | | } |
+361 | |
362 | 152 488 | | | | 150 226 | @identities = ( sort {$b <=> $a} @identities ); # sort descending numerically |
+363 | 152 | | | | 282 | return (@identities); # return sorted identity hits |
+364 | | | | | | } |
+
+
+
diff --git a/cover_db/cover.14 b/cover_db/cover.14
new file mode 100644
index 0000000..d8a5875
--- /dev/null
+++ b/cover_db/cover.14
@@ -0,0 +1 @@
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\ No newline at end of file
diff --git a/cover_db/cover.css b/cover_db/cover.css
new file mode 100644
index 0000000..d875957
--- /dev/null
+++ b/cover_db/cover.css
@@ -0,0 +1,75 @@
+/* Stylesheet for Devel::Cover HTML reports */
+
+/* You may modify this file to alter the appearance of your coverage
+ * reports. If you do, you should probably flag it read-only to prevent
+ * future runs from overwriting it.
+ */
+
+/* Note: default values use the color-safe web palette. */
+
+body {
+ font-family: sans-serif;
+}
+
+h1 {
+ background-color: #3399ff;
+ border: solid 1px #999999;
+ padding: 0.2em;
+ -moz-border-radius: 10px;
+}
+
+a {
+ color: #000000;
+}
+a:visited {
+ color: #333333;
+}
+
+table {
+ border-collapse: collapse;
+ border-spacing: 0px;
+}
+tr {
+ text-align : center;
+ vertical-align: top;
+}
+th,.h {
+ background-color: #cccccc;
+ border: solid 1px #333333;
+ padding: 0em 0.2em;
+}
+td {
+ border: solid 1px #cccccc;
+}
+
+/* source code */
+pre,.s {
+ text-align: left;
+ font-family: monospace;
+ white-space: pre;
+ padding: 0em 0.5em 0em 0.5em;
+}
+
+/* Classes for color-coding coverage information:
+ * c0 : path not covered or coverage < 75%
+ * c1 : coverage >= 75%
+ * c2 : coverage >= 90%
+ * c3 : path covered or coverage = 100%
+ */
+.c0, .c1, .c2, .c3 { text-align: right; }
+.c0 {
+ background-color: #ff9999;
+ border: solid 1px #cc0000;
+}
+.c1 {
+ background-color: #ffcc99;
+ border: solid 1px #ff9933;
+}
+.c2 {
+ background-color: #ffff99;
+ border: solid 1px #cccc66;
+}
+.c3 {
+ background-color: #99ff99;
+ border: solid 1px #009900;
+}
diff --git a/cover_db/coverage.html b/cover_db/coverage.html
new file mode 100644
index 0000000..d7629b3
--- /dev/null
+++ b/cover_db/coverage.html
@@ -0,0 +1,41 @@
+
+
+
+
+
+line | stmt | bran | cond | sub | time | code |
+1 | | | | | | package Search; |
+2 | |
3 | 140 140 140 | | | | 435 131 7277 | use Exporter qw(import); |
+4 | | | | | | our @ISA = qw(Exporter); |
+5 | | | | | | our @EXPORT = qw(findOligo); #functions exported by default |
+6 | | | | | | our @EXPORT_OK = qw(blast); #functions for explicit export |
+7 | |
8 | 140 140 140 140 140 140 140 140 140 140 140 140 | | | | 399 119 2741 329 117 2364 310 94 479 2388 116 5492 | use strict; use warnings; use diagnostics; use feature qw(say); |
+9 | 140 140 140 | | | | 320 904 4571 | use Carp; |
+10 | |
11 | 140 140 140 140 140 140 | | | | 125022 8694531 4130 86328 2663478 3458 | use Bio::Seq; use Bio::SeqIO; |
+12 | 140 140 140 | | | | 123942 6795200 1434 | use Bio::Tools::Run::StandAloneBlastPlus; |
+13 | 140 140 140 140 140 140 | | | | 3283 160 4714 391 120 102564 | use MyConfig; use MyIO; |
+14 | |
15 | |
16 | | | | | | # ============================================================================== |
+17 | | | | | | # |
+18 | | | | | | # CAPITAN: Andres Breton, http://andresbreton.com |
+19 | | | | | | # FILE: Search.pm |
+20 | | | | | | # LICENSE: |
+21 | | | | | | # USAGE: |
+22 | | | | | | # DEPENDENCIES: - NCBI's BLAST+ CL utility |
+23 | | | | | | # |
+24 | | | | | | # ============================================================================== |
+25 | |
26 - 52 | | =head1 NAME
+
+Search - package searching gRNA sequences and offsite targets for CRISPR targeting
+
+=head1 SYNOPSIS
+
+Creation:
+ use Search;
+
+=head1 DESCRIPTION
+
+
+=head1 EXPORTS
+
+=head2 Default Behaviors
+
+Exports findOligo subroutine by default
+
+use Search;
+
+=head2 Optional Behaviors
+
+Search::blast;
+
+=head1 FUNCTIONS
+
+=cut |
+53 | |
54 | | | | | | #------------------------------------------------------------------------------- |
+55 | | | | | | # MAIN |
+56 | |
57 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+58 - 72 | | =head2 findOligo
+
+ Arg [1] : Sequence to be searched details hash
+
+ Arg [2] : Window size of gRNA sequence
+
+ Example : findOligo($seqDetails, $windowSize)
+
+ Description : Find gRNA sequences for CRISPR targeting
+
+ Returntype : Hash of hashes reference
+
+ Status : Stable
+
+=cut |
+73 | | | | | | sub findOligo { |
+74 | 802 | | | | 1984 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '($seqDetails, $windowSize)'; |
+75 | 802 | | | | 35379 | @_ == 2 or confess wrongNumberArguments(), $filledUsage; |
+76 | |
77 | 472 | | | | 699 | my ($seqDetails, $windowSize) = @_; |
+78 | 472 | | | | 441 | my %gRNAs; |
+79 | 472 | | | | 456 | my $instance = 0; # track gRNA count |
+80 | |
81 | | | | | | # Anonymous subroutine to find gRNA sequences in forward and reverse strands. |
+82 | | | | | | # Stores both strand findings in same hash (%gRNAs) containing all information |
+83 | | | | | | my $go = sub { |
+84 | 472 | | | | 799 | my ($sequence, $strand) = @_; |
+85 | |
86 | 472 | | | | 5041 | say "Searching gRNA sequences on $strand strand"; |
+87 | |
88 | 472 | | | | 647 | my $seqLen = length($sequence); |
+89 | 472 | | | | 559 | my ($kmer, $PAM, $gRNA, $content, $contentG, $contentC, $GC); |
+90 | |
91 | 472 | | | | 4333 | for (my $i = 0; $i < $seqLen; $i++) { |
+92 | 98648 | | | | 52106 | my $window = substr $sequence, $i, $windowSize; |
+93 | |
94 | | | | | | # LAST STEP: When DONE LOOKING UP -- Return gRNA sequences and information. |
+95 | | | | | | # Returns references of HoH containing all gRNA instances found and |
+96 | | | | | | # respective information for each gRNA sequence |
+97 | 98648 | | | | 81239 | return(\%gRNAs) if ( length($window) < $windowSize ); # don't go out of bounds when at end of sequence |
+98 | |
99 | 98176 | | | | 144898 | if ($window =~ /(.+)(.GG)$/) { |
+100 | 4012 | | | | 6544 | ($kmer, $PAM) = ($1, $2); # get first 'kmer' and PAM nucleotides in gRNA (kmer + PAM) |
+101 | 4012 4012 | | | | 4100 2428 | my $name = "gRNA_$instance"; $instance++; |
+102 | 4012 | | | | 2490 | my $gRNA = $kmer . $PAM; # kmer + PAM = gRNA sequence |
+103 | 4012 | | | | 4217 | my $palindrome = _palindrome($kmer); |
+104 | |
105 | | | | | | # GC Content |
+106 | 4012 | | | | 3187 | $contentG = $gRNA =~ tr/G//; |
+107 | 4012 | | | | 2667 | $contentC = $gRNA =~ tr/C//; |
+108 | 4012 | | | | 3259 | $GC = ($contentG + $contentC)/$windowSize; |
+109 | |
110 | | | | | | # Store gRNA sequences and info in Hash of Hashes |
+111 | 4012 | | | | 16074 | $content = { # anonymous hash of relevant gRNA content |
+112 | | | | | | 'kmer' => $kmer, |
+113 | | | | | | 'palindrome' => $palindrome, |
+114 | | | | | | 'strand' => $strand, |
+115 | | | | | | 'start' => $i, |
+116 | | | | | | 'sequence' => $gRNA, |
+117 | | | | | | 'PAM' => $PAM, |
+118 | | | | | | 'G' => $contentG, |
+119 | | | | | | 'C' => $contentC, |
+120 | | | | | | 'GC' => $GC, |
+121 | | | | | | }; |
+122 | | | | | | # Hash key == gRNA sequence name |
+123 | | | | | | # Hash value == Hash with gRNA content info |
+124 | 4012 | | | | 9151 | $gRNAs{$name} = $content; |
+125 | | | | | | } |
+126 | | | | | | } |
+127 | 472 | | | | 4267 | }; |
+128 | |
129 | | | | | | # Get all gRNA sequences in forward and reverse strands of sequence passed, -seq |
+130 | 472 | | | | 3290 | $go->( $seqDetails->{'sequence'}, 'plus' ); |
+131 | 236 | | | | 791 | $go->( $seqDetails->{'reverse'}, 'reverse' ); |
+132 | | | | | | } |
+133 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+134 - 150 | | =head2 blast
+
+ Arg [1] : gRNA fasta file
+
+ Arg [2] : Array reference with subject sequence file(s) provided for search
+
+ Arg [3] : Output file name
+
+ Example : Search::blast($fastaFile, \@SUBJSEQS, $OUTFILE);
+
+ Description : Run BLAST+ search for gRNA sequences
+
+ Returntype : Hash reference
+
+ Status : Stable
+
+=cut |
+151 | | | | | | sub blast { |
+152 | 880 | | | | 2725 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '(\%fastaFile, \@SUBJSEQS, $OUTFILE)'; |
+153 | 880 | | | | 37927 | @_ == 3 or confess wrongNumberArguments(), $filledUsage; |
+154 | |
155 | 538 | | | | 1204 | my ($fasta, $SUBJSEQS, $OUTFILE) = @_; |
+156 | 538 | | | | 3550 | my @SUBJSEQS = @$SUBJSEQS; |
+157 | 196 | | | | 297 | my (%targets, $info, $hsps); |
+158 | 196 | | | | 364 | my $outDir = $main::OUTDIR; |
+159 | 196 | | | | 283 | my $wordSize = 7; |
+160 | |
161 | 196 | | | | 1553 | mkDir("$outDir/blast"); |
+162 | |
163 | 196 | | | | 118277 | foreach my $subject (@SUBJSEQS) { |
+164 | 154 | | | | 555 | my $subjName = _getSeqName($subject); |
+165 | 154 | | | | 14652 | my $outFile = "$outDir/blast/$subjName\_$OUTFILE\_blast.txt"; |
+166 | |
167 | 154 | | | | 1941 | say "Searching gRNA sequences against $subject"; |
+168 | |
169 | | | | | | # Create StandAloneBlastPlus Factory |
+170 | 154 | | | | 1999 | my $fac = Bio::Tools::Run::StandAloneBlastPlus->new( |
+171 | | | | | | -db_data => $subject, |
+172 | | | | | | ); |
+173 | |
174 | | | | | | # Perform BLAST call |
+175 | 154 | | | | 170900 | $fac->blastn( -query => $fasta, |
+176 | | | | | | -outfile => $outFile, |
+177 | | | | | | -method_args => [ -word_size => 7], |
+178 | | | | | | ); |
+179 | |
180 | | | | | | # Rewind to beginning of results and get all |
+181 | 154 | | | | 32674662 | $fac->rewind_results; |
+182 | |
183 | | | | | | # Process each gRNA sequence |
+184 | 154 | | | | 334139 | while ( my $result = $fac->next_result ) { |
+185 | 2618 | | | | 11403916 | my ($gRNA) = $result->query_name =~ /(.*):\d+/;; # gRNA sequence name ex.) 'gRNA_0', removes appendend positioning |
+186 | 2618 | | | | 26230 | my $numHits = $result->num_hits; |
+187 | |
188 | | | | | | # Resolve when gRNA sequences has no matches in subject |
+189 | 2618 | | | | 712400 | say "\t$gRNA has $numHits hits here" if ($numHits == 0); |
+190 | |
191 | | | | | | # Default anonymous hashes for storing BLAST results |
+192 | | | | | | # $info hash stores each gRNA BLAST result's info |
+193 | | | | | | # $hsps hash stores that match's High-scoring Segment Pairs (HSPs) result |
+194 | 2618 | | | | 6829 | $info = { |
+195 | | | | | | 'numhits' => $numHits, |
+196 | | | | | | 'occurrences' => 0, |
+197 | | | | | | }; |
+198 | |
199 | 2618 | | | | 13686 | $hsps = { |
+200 | | | | | | 'rank' => 0, |
+201 | | | | | | 'qstart' => 0, |
+202 | | | | | | 'qend' => 0, |
+203 | | | | | | 'sstart' => 0, |
+204 | | | | | | 'send' => 0, |
+205 | | | | | | 'sstrand' => 0, |
+206 | | | | | | 'pident' => 0, |
+207 | | | | | | 'nident' => 0, |
+208 | | | | | | 'gaps' => 0, |
+209 | | | | | | }; |
+210 | |
211 | | | | | | # Process each gRNA hit |
+212 | 2618 | | | | 5418 | while ( my $hit = $result->next_hit ) { |
+213 | |
214 | 2618 | | | | 134395 | $info->{'occurrences'} = $hit->num_hsps; |
+215 | |
216 | | | | | | # Process each match (HSP) in iterative fashion |
+217 | 2618 | | | | 13783 | while( my $hsp = $hit->next_hsp ) { |
+218 | | | | | | # Get all values to store in hashes |
+219 | 9394 | | | | 4292562 | $hsps = { |
+220 | | | | | | 'rank' => $hsp->rank, |
+221 | | | | | | 'qstart' => $hsp->start('query'), |
+222 | | | | | | 'qend' => $hsp->end('query'), |
+223 | | | | | | 'qstrand' => $hsp->strand('query'), |
+224 | | | | | | 'sstart' => $hsp->start('hit'), |
+225 | | | | | | 'send' => $hsp->end('hit'), |
+226 | | | | | | 'sstrand' => $hsp->strand('hit'), |
+227 | | | | | | 'pident' => $hsp->percent_identity, |
+228 | | | | | | 'nident' => $hsp->num_identical, |
+229 | | | | | | 'gaps' => $hsp->gaps, |
+230 | | | | | | }; |
+231 | 9394 9394 | | | | 10608404 48657 | push @{ $targets{$gRNA}{$subjName}{'hsps'} } , $hsps; |
+232 | | | | | | } |
+233 | | | | | | } |
+234 | 2618 2618 | | | | 76153 15630 | push @{ $targets{$gRNA}{$subjName}{'info'} } , $info; |
+235 | | | | | | } |
+236 | 154 | | | | 61504 | $fac->cleanup; #clean up temp database files |
+237 | | | | | | } |
+238 | |
239 | 196 | | | | 206889 | say "\nBLAST files saved in: '$outDir/blast' "; |
+240 | |
241 | | | | | | # Hash of Hashes of Hashes of Arrays of Hash to store BLAST results for each query |
+242 | | | | | | # -- Hash key == gRNA name |
+243 | | | | | | # -- Hash key == Subject name |
+244 | | | | | | # -- Hash key == 'info' |
+245 | | | | | | # -- Hash key == 'hsps' |
+246 | | | | | | # -- Array accounts for multiple hits for each gRNA sequence as hashes.... |
+247 | | | | | | # -- Hash contains BLAST match info |
+248 | 196 | | | | 2793 | return(\%targets); |
+249 | | | | | | } |
+250 | |
251 | | | | | | #------------------------------------------------------------------------------- |
+252 | | | | | | # HELPERS |
+253 | |
254 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+255 | | | | | | # $input = ($seq); |
+256 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+257 | | | | | | # This function takes 1 argument, a record file and extracts relevant information |
+258 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+259 | | | | | | # $return = ($name); Name of sequence record |
+260 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+261 | | | | | | sub _getSeqName { |
+262 | 154 | | | | 462 | my ($seq) = @_; |
+263 | |
264 | | | | | | # Sequence OO |
+265 | 154 | | | | 2700 | my $seqInObj = Bio::SeqIO->new(-file => $seq, -alphabet => "dna"); |
+266 | 154 | | | | 102705 | my $format = $seqInObj->_guess_format($seq); #check format of input file |
+267 | 154 | | | | 2326 | my $seqObj = $seqInObj->next_seq; |
+268 | 154 | | | | 18261 | my $name = $seqObj->display_id; |
+269 | |
270 | 154 | | | | 3074 | return $name; |
+271 | | | | | | } |
+272 | |
273 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+274 | | | | | | # $input = ($sequence); |
+275 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+276 | | | | | | # This function takes 1 argument, a sequence string and determines if string is |
+277 | | | | | | # palindrome. |
+278 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+279 | | | | | | # $return = Returns yes/no if palindrome found |
+280 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+281 | |
282 | | | | | | sub _palindrome { |
+283 | 4440 | | | | 6832 | my $filledUsage = 'Usage: ' . (caller(0))[3] . '($sequence)'; |
+284 | 4440 | | | | 117569 | @_ == 1 or confess wrongNumberArguments(), $filledUsage; |
+285 | |
286 | 4333 | | | | 3645 | my ($sequence) = @_; |
+287 | |
288 | 4333 | | | | 3161 | my $palindrome = reverse $sequence; |
+289 | |
290 | 4333 | | | | 8436 | $palindrome eq $sequence ? return("Yes") : return("No"); |
+291 | | | | | | } |
+292 | |
293 | |
294 | |
295 - 313 | | =head1 COPYRIGHT AND LICENSE
+
+Andres Breton ©
+
+[LICENSE]
+
+=head1 CONTACT
+
+Please email comments or questions to Andres Breton, <dev@andresbreton.com>
+
+=head1 SETTING PATH
+
+If PERL5LIB was not set, do something like this:
+
+use FindBin; use lib "$FindBin::RealBin/lib";
+
+This finds and uses subdirectory 'lib' in current directoy as library location
+
+=cut |
+314 | | | | | | 1; |
+
+
+
diff --git a/cover_db/runs/1490301145.52861.39223/cover.14 b/cover_db/runs/1490301145.52861.39223/cover.14
deleted file mode 100644
index 25c78a7..0000000
--- a/cover_db/runs/1490301145.52861.39223/cover.14
+++ /dev/null
@@ -1 +0,0 @@
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\ No newline at end of file
diff --git a/cover_db/structure/0bf19b40fe954a61ff714e5a05d832e5 b/cover_db/structure/0bf19b40fe954a61ff714e5a05d832e5
index 9568ce6..213590b 100644
--- a/cover_db/structure/0bf19b40fe954a61ff714e5a05d832e5
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diff --git a/cover_db/structure/e6845289bc529cbfc889f530bf091645 b/cover_db/structure/e6845289bc529cbfc889f530bf091645
index 02519d9..6cc738d 100644
--- a/cover_db/structure/e6845289bc529cbfc889f530bf091645
+++ b/cover_db/structure/e6845289bc529cbfc889f530bf091645
@@ -1 +1 @@
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diff --git a/cover_db/t-chomp-t--branch.html b/cover_db/t-chomp-t--branch.html
new file mode 100644
index 0000000..e1e1715
--- /dev/null
+++ b/cover_db/t-chomp-t--branch.html
@@ -0,0 +1,31 @@
+
+
+
+
+
+line | stmt | bran | cond | sub | time | code |
+1 | | | | | | #!/usr/bin/env perl |
+2 | |
3 | 1 1 1 1 1 1 1 1 1 1 1 1 | | | | 16785 4 30 3 0 15 556 82807 5 34 1 50 | use strict; use warnings; use diagnostics; use feature qw(say); |
+4 | |
5 | 1 1 1 | | | | 1606 26512 5 | use IO::Detect qw(is_filehandle); |
+6 | 1 1 1 | | | | 1450 41293 6 | use Test::More tests => 73; |
+7 | 1 1 1 | | | | 1331 1952 3 | use Test::Exception; #need this to get dies_ok and lives_ok to work |
+8 | |
9 | 1 1 1 1 1 1 | | | | 417 578 30 263 339 3 | use FindBin; use lib "$FindBin::RealBin/../lib"; |
+10 | |
11 | 1 | | | | 108243 | my $SEQ = 'usr/test.fasta'; |
+12 | 1 | | | | 2 | my @SUBJSEQS = $SEQ; |
+13 | 1 | | | | 28 | my $OUTFILE = 'test'; |
+14 | 1 | | | | 2 | our $OUTDIR = 'tests'; |
+15 | 1 | | | | 1 | our $WINDOWSIZE = 23; |
+16 | 1 | | | | 1 | my $VERBOSE; |
+17 | 1 | | | | 1 | my $SS; |
+18 | 1 | | | | 1 | our $HTML; |
+19 | 1 | | | | 1 | my $USAGE = "Usage message"; |
+20 | |
21 | 1 | | | | 5 | mkDir($OUTDIR); |
+22 | |
23 | | | | | | #------------------------------------------------------------------------------- |
+24 | | | | | | # TESTS |
+25 | |
26 | 1 | | | | 119 | require_ok 'chomp.pl'; |
+27 | |
28 | | | | | | # Modules |
+29 | 1 1 1 1 1 | | | | 113 508 4424 1 61 | BEGIN { use_ok('MyIO') } |
+30 | 1 1 1 1 1 | | | | 226 87 1 0 25 | BEGIN { use_ok('MyConfig') } |
+31 | 1 1 1 1 1 | | | | 195 576 300 1 13 | BEGIN { use_ok('Handlers') } |
+32 | 1 1 1 1 1 | | | | 183 393 2 1 21 | BEGIN { use_ok('Search') } |
+33 | 1 1 1 1 1 | | | | 228 438 3 1 23 | BEGIN { use_ok('SS') } |
+34 | |
35 | |
36 | | | | | | #- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
+37 | | | | | | # MAIN |
+38 | |
39 | | | | | | #------------------------------------------------------------------------------- |
+40 | | | | | | # checks |
+41 | 1 | | | | 268 | print "\nTesting checks()...\n"; |
+42 | 1 1 | | | | 5 17 | dies_ok { checks() } 'dies ok when no arguments passed in checks()'; |
+43 | |
44 | | | | | | #------------------------------------------------------------------------------- |
+45 | | | | | | # setParameters |
+46 | | | | | | # print "\nTesting setParameters()...\n"; |
+47 | 1 1 | | | | 755 45 | dies_ok { setParameters() } 'dies ok when no arguments passed in setParameters()'; |
+48 | |
49 | | | | | | #------------------------------------------------------------------------------- |
+50 | | | | | | # getSeqDetails |
+51 | 1 | | | | 218 | print "\nTesting getSeqDetails()...\n"; |
+52 | 1 1 | | | | 5 34 | dies_ok { getSeqDetails() } 'dies ok when no argument passed in getSeqDetails()'; |
+53 | 1 1 | | | | 358 21 | dies_ok { getSeqDetails(1) } 'dies ok when 1 incorrect argument is passed in getSeqDetails()'; |
+54 | 1 1 | | | | 1617 17 | dies_ok { getSeqDetails(1,2) } 'dies ok when >1 arguments are passed in getSeqDetails()'; |
+55 | 1 1 | | | | 257 18 | dies_ok { getSeqDetails("not.fasta") } 'dies ok when argument is not FASTA file in getSeqDetails()'; |
+56 | 1 1 | | | | 4811 16 | lives_ok { getSeqDetails($SEQ) } 'lives ok when file is passed in getSeqDetails()'; |
+57 | 1 | | | | 475 | my ($seqDetails) = getSeqDetails($SEQ); |
+58 | |
59 | | | | | | #------------------------------------------------------------------------------- |
+60 | | | | | | # findOligo() |
+61 | 1 | | | | 16 | print "\nTesting findOligo()...\n"; |
+62 | 1 1 | | | | 9 32 | dies_ok { findOligo() } 'dies ok when no argument passed in findOligo()'; |
+63 | 1 1 | | | | 690 13 | dies_ok { findOligo(1) } 'dies ok when 1 argument is passed in findOligo()'; |
+64 | 1 1 | | | | 585 16 | dies_ok { findOligo(1, $WINDOWSIZE) } 'dies ok when 2 incorrect arguments passed in findOligo()'; |
+65 | 1 1 | | | | 173 12 | dies_ok { findOligo(1, $WINDOWSIZE) } 'dies ok when 2 arguments passed, but 1 is incorrect in findOligo()'; |
+66 | 1 1 | | | | 205 36 | dies_ok { findOligo(1,2,3) } 'dies ok when 3 arguments are passed in findOligo()'; |
+67 | 1 1 | | | | 546 12 | lives_ok { findOligo($seqDetails, $WINDOWSIZE) } 'lives ok when right paramaterers passed in findOligo()'; |
+68 | |
69 | 1 | | | | 212 | my ($gRNAs, $CRPseqs) = findOligo($seqDetails, $WINDOWSIZE); |
+70 | 1 | | | | 5 | is(ref($gRNAs), "HASH", "gRNA hash reference returned"); |
+71 | 1 | | | | 406 | is($gRNAs->{'gRNA_0'}->{'sequence'}, 'ATGTAGCTAGCTAGCTAGTAGGG', 'gRNA_0 sequence is correct'); |
+72 | 1 | | | | 177 | is($gRNAs->{'gRNA_1'}->{'sequence'}, 'AAAAAATTTTCTCTATCTAACGG', 'gRNA_1 sequence is correct'); |
+73 | 1 | | | | 158 | is($gRNAs->{'gRNA_2'}->{'sequence'}, 'AAAAATTTTCTCTATCTAACGGG', 'gRNA_2 sequence is correct'); |
+74 | 1 | | | | 149 | is($gRNAs->{'gRNA_3'}->{'sequence'}, 'TGTGATCACGTACTATTATGCGG', 'gRNA_3 sequence is correct'); |
+75 | 1 | | | | 133 | is($gRNAs->{'gRNA_4'}->{'sequence'}, 'AAAAATCCCATCGATCTAGCAGG', 'gRNA_4 sequence is correct'); |
+76 | 1 | | | | 150 | is($gRNAs->{'gRNA_5'}->{'sequence'}, 'TCCCATCGATCTAGCAGGCCCGG', 'gRNA_5 sequence is correct'); |
+77 | 1 | | | | 133 | is($gRNAs->{'gRNA_6'}->{'sequence'}, 'CGAAAAAAAATTTTCCCTATCGG', 'gRNA_6 sequence is correct'); |
+78 | 1 | | | | 147 | is($gRNAs->{'gRNA_7'}->{'sequence'}, 'GAAAAAAAATTTTCCCTATCGGG', 'gRNA_7 sequence is correct'); |
+79 | 1 | | | | 131 | is($gRNAs->{'gRNA_8'}->{'sequence'}, 'AAAAAAAATTTTCCCTATCGGGG', 'gRNA_8 sequence is correct'); |
+80 | 1 | | | | 150 | is($gRNAs->{'gRNA_9'}->{'sequence'}, 'AAAAAAATTTTCCCTATCGGGGG', 'gRNA_9 sequence is correct'); |
+81 | 1 | | | | 130 | is($gRNAs->{'gRNA_10'}->{'sequence'}, 'CTATAGCATGGGCCCCCGATAGG', 'gRNA_10 sequence is correct'); |
+82 | 1 | | | | 151 | is($gRNAs->{'gRNA_11'}->{'sequence'}, 'TATAGCATGGGCCCCCGATAGGG', 'gRNA_11 sequence is correct'); |
+83 | 1 | | | | 131 | is($gRNAs->{'gRNA_12'}->{'sequence'}, 'TTCGTCATGCATGCTCGCTCCGG', 'gRNA_12 sequence is correct'); |
+84 | 1 | | | | 148 | is($gRNAs->{'gRNA_13'}->{'sequence'}, 'TCGTCATGCATGCTCGCTCCGGG', 'gRNA_13 sequence is correct'); |
+85 | 1 | | | | 134 | is($gRNAs->{'gRNA_14'}->{'sequence'}, 'CTCCGGGCCTGCTAGATCGATGG', 'gRNA_14 sequence is correct'); |
+86 | 1 | | | | 150 | is($gRNAs->{'gRNA_15'}->{'sequence'}, 'TCCGGGCCTGCTAGATCGATGGG', 'gRNA_15 sequence is correct'); |
+87 | 1 | | | | 130 | is($gRNAs->{'gRNA_16'}->{'sequence'}, 'ATAGTACGTGATCACAGTCATGG', 'gRNA_16 sequence is correct'); |
+88 | |
89 | | | | | | #------------------------------------------------------------------------------- |
+90 | | | | | | # writeFasta |
+91 | 1 | | | | 153 | print "\nTesting writeFasta()...\n"; |
+92 | 1 1 | | | | 7 25 | dies_ok { writeFasta() } 'dies ok when no argument passed in writeFasta()'; |
+93 | 1 1 | | | | 263 39 | dies_ok { writeFasta(1) } 'dies ok when 1 argument is passed in writeFasta()'; |
+94 | 1 1 | | | | 193 13 | dies_ok { writeFasta(1,2) } 'dies ok when 2 incorrect arguments passed in writeFasta()'; |
+95 | 1 1 | | | | 201 22 | dies_ok { writeFasta(1, $OUTFILE) } 'dies ok when 2 arguments passed, but 1 is incorrect in writeFasta()'; |
+96 | 1 1 | | | | 206 17 | dies_ok { writeFasta(1,2,3) } 'dies ok when 3 arguments are passed in writeFasta()'; |
+97 | 1 1 | | | | 229 20 | lives_ok { writeFasta($gRNAs, $OUTFILE) } 'lives ok when right parameters are passed in writeFasta()'; |
+98 | 1 | | | | 352 | my $fastaFile = writeFasta($gRNAs, $OUTFILE); |
+99 | |
100 | | | | | | #------------------------------------------------------------------------------- |
+101 | | | | | | # ss |
+102 | 1 | | | | 6 | mkDir("$OUTDIR/ss"); |
+103 | 1 1 | | | | 79 21 | dies_ok { ss() } 'dies ok when no argument passed in ss()'; |
+104 | 1 1 | | | | 635 10 | dies_ok { ss(1) } 'dies ok when 1 argument passed in ss()'; |
+105 | 1 1 | | | | 703 12 | dies_ok { ss(1,2) } 'dies ok when 2 invalid arguments passed in ss()'; |
+106 | 1 1 | | | | 285 19 | lives_ok { ss('gRNA_0', $gRNAs->{'gRNA_0'}->{'sequence'} ) } 'lives ok when correct arguments passed in ss()'; |
+107 | |
108 | | | | | | #------------------------------------------------------------------------------- |
+109 | | | | | | # Search::blast |
+110 | 1 | | | | 740 | print "\nTesting Search::blast()...\n"; |
+111 | 1 1 | | | | 63 63 | dies_ok { Search::blast() } 'dies ok when no argument passed in Search::blast()'; |
+112 | 1 1 | | | | 878 18 | dies_ok { Search::blast(1) } 'dies ok when 1 argument is passed in Search::blast()'; |
+113 | 1 1 | | | | 611 26 | dies_ok { Search::blast(1,2,) } 'dies ok when 2 arguments passed, but at least 1 is incorrect in Search::blast()'; |
+114 | 1 1 | | | | 573 14 | dies_ok { Search::blast(1,2, $OUTFILE) } 'dies ok when 3 arguments passed, but 1 is incorrect in Search::blast()'; |
+115 | 1 1 | | | | 182 23 | dies_ok { Search::blast(1, $fastaFile, $OUTFILE) } 'dies ok when 3 arguments passed, but 2 are incorrect in Search::blast()'; |
+116 | 1 1 | | | | 171 21 | dies_ok { Search::blast(1,2,3) } 'dies ok when 3 incorrect arguments are passed in Search::blast()'; |
+117 | 1 1 | | | | 178 22 | lives_ok { Search::blast($fastaFile, \@SUBJSEQS, $OUTFILE) } 'lives ok when right parameters are passed in Search::blast()'; |
+118 | 1 | | | | 762 | my $targets = Search::blast($fastaFile, \@SUBJSEQS, $OUTFILE); |
+119 | |
120 | | | | | | #------------------------------------------------------------------------------- |
+121 | | | | | | # _palindrome |
+122 | 1 | | | | 6 | print "\nTesting Search::_palindrome()...\n"; |
+123 | 1 1 | | | | 26 50 | dies_ok { Search::_palindrome() } 'dies ok when no argument passed in Search::_palindrome()'; |
+124 | | | | | | # dies_ok { Search::_palindrome(1) } 'dies ok when argument passed is incorrect in Search::_palindrome()'; |
+125 | 1 1 | | | | 938 22 | lives_ok { Search::_palindrome('ATGTA') } 'lives ok when right parameters are passed in Search::_palindrome()'; |
+126 | 1 | | | | 225 | is ( Search::_palindrome('ATGTA') , 'Yes', 'YES palindrome found is correct'); |
+127 | 1 | | | | 235 | is ( Search::_palindrome('ATGTAA') , 'No', 'NO palindrome found is correct'); |
+128 | |
129 | | | | | | #------------------------------------------------------------------------------- |
+130 | | | | | | # writeResults |
+131 | 1 | | | | 267 | print "\nTesting writeResults()...\n"; |
+132 | 1 1 | | | | 10 42 | dies_ok { writeResults() } 'dies ok when no argument passed in writeResults()'; |
+133 | 1 1 | | | | 318 22 | dies_ok { writeResults(1) } 'dies ok when 1 argument is passed in writeResults()'; |
+134 | 1 1 | | | | 271 23 | dies_ok { writeResults(1,2) } 'dies ok when 2 incorrect arguments passed in writeResults()'; |
+135 | 1 1 | | | | 310 19 | dies_ok { writeResults(1,2,3) } 'dies ok when 3 incorrect arguments passed in writeResults()'; |
+136 | 1 1 | | | | 218 12 | dies_ok { writeResults(1,2,3,4) } 'dies ok when 4 incorrect arguments passed in writeResults()'; |
+137 | 1 1 | | | | 250 17 | dies_ok { writeResults(1,2,3,4,5) } 'dies ok when 5 incorrect arguments passed in writeResults()'; |
+138 | 1 1 | | | | 237 16 | dies_ok { writeResults(1,2,3,4,5,6) } 'dies ok when 6 incorrect arguments passed in writeResults()'; |
+139 | |
140 | 1 | | | | 169 | my $DOWNSEQ; |
+141 | 1 | | | | 1 | my $UPSEQ; |
+142 | 1 1 | | | | 13 14 | lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok whith both undef DOWN/UP-SEQ parameters are passed in writeResults()'; |
+143 | |
144 | 1 | | | | 314 | $DOWNSEQ = "DWDWDW"; |
+145 | 1 1 | | | | 8 31 | lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok whith undef UPSEQ parameter is passed in writeResults()'; |
+146 | |
147 | 1 | | | | 320 | $DOWNSEQ = ""; |
+148 | 1 | | | | 2 | $UPSEQ = "UPUPUP"; |
+149 | 1 1 | | | | 48 24 | lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok whith undef DOWNSEQ & defined $UPSEQ parameter is passed in writeResults()'; |
+150 | |
151 | |
152 | 1 | | | | 262 | $DOWNSEQ = "DWDWDW"; |
+153 | 1 | | | | 3 | $UPSEQ = "UPUPUP"; |
+154 | 1 1 | | | | 26 22 | lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok when right parameters are passed in writeResults()'; |
+155 | 1 | | | | 287 | writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE); |
+156 | |
157 | | | | | | #------------------------------------------------------------------------------- |
+158 | | | | | | # sortIdentities |
+159 | 1 | | | | 4 | print "\nTesting sortIdentities()...\n"; |
+160 | 1 1 | | | | 25 31 | dies_ok { sortIdentities() } 'dies ok when no arguments passed'; |
+161 | 1 1 | | | | 317 15 | dies_ok { sortIdentities(1) } 'dies ok when invalid argument passed'; |
+162 | 1 1 | | | | 184 17 | dies_ok { sortIdentities(1,2) } 'dies ok when 2 arguments passed'; |
+163 | 1 1 | | | | 198 13 | lives_ok { sortIdentities( $targets->{'gRNA_0'}{'test'}{'hsps'} ) } 'lives ok when 1 correct arguments passed'; |
+164 | |
165 | |
166 | | | | | | #------------------------------------------------------------------------------- |
+167 | | | | | | # Clean Up |
+168 | 1 | | | | 4770 | `rm -r ./$OUTDIR`; |
+169 | |
170 | 1 | | | | 41 | done_testing(); |
+
+
+
diff --git a/t/chomp.t b/t/chomp.t
index d06b8de..f06c943 100644
--- a/t/chomp.t
+++ b/t/chomp.t
@@ -3,22 +3,20 @@
use strict; use warnings; use diagnostics; use feature qw(say);
use IO::Detect qw(is_filehandle);
-use Test::More;
+use Test::More tests => 73;
use Test::Exception; #need this to get dies_ok and lives_ok to work
use FindBin; use lib "$FindBin::RealBin/../lib";
-my $SEQ = "usr/test.fasta";
-my @GENOME;
-my $OUTFILE = "test";;
-my $DOWNSEQ = "DWDWDW";
-my $UPSEQ = "UPUPUP";
+my $SEQ = 'usr/test.fasta';
+my @SUBJSEQS = $SEQ;
+my $OUTFILE = 'test';
our $OUTDIR = 'tests';
our $WINDOWSIZE = 23;
my $VERBOSE;
-my $SS = "";
-our $HTML = "";
-my @SUBJSEQS;
+my $SS;
+our $HTML;
+my $USAGE = "Usage message";
mkDir($OUTDIR);
@@ -38,18 +36,17 @@ BEGIN { use_ok('SS') }
#- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# MAIN
-# # checks
-# print "\nTesting checks()...\n";
-# dies_ok { checks() } 'dies ok when no arguments passed in checks()';
-#
-#
-# # setParameters
-# print "\nTesting setParameters()...\n";
-# # dies_ok { setParameters(1) } 'dies ok when 1 arguments is passed in setParameters()';
-# dies_ok { setParameters() } 'dies ok when no arguments passed in setParameters()';
-# # setParameters();
+#-------------------------------------------------------------------------------
+# checks
+print "\nTesting checks()...\n";
+dies_ok { checks() } 'dies ok when no arguments passed in checks()';
+#-------------------------------------------------------------------------------
+# setParameters
+# print "\nTesting setParameters()...\n";
+dies_ok { setParameters() } 'dies ok when no arguments passed in setParameters()';
+#-------------------------------------------------------------------------------
# getSeqDetails
print "\nTesting getSeqDetails()...\n";
dies_ok { getSeqDetails() } 'dies ok when no argument passed in getSeqDetails()';
@@ -59,14 +56,14 @@ dies_ok { getSeqDetails("not.fasta") } 'dies ok when argument is not FASTA file
lives_ok { getSeqDetails($SEQ) } 'lives ok when file is passed in getSeqDetails()';
my ($seqDetails) = getSeqDetails($SEQ);
-
+#-------------------------------------------------------------------------------
# findOligo()
print "\nTesting findOligo()...\n";
dies_ok { findOligo() } 'dies ok when no argument passed in findOligo()';
dies_ok { findOligo(1) } 'dies ok when 1 argument is passed in findOligo()';
dies_ok { findOligo(1, $WINDOWSIZE) } 'dies ok when 2 incorrect arguments passed in findOligo()';
dies_ok { findOligo(1, $WINDOWSIZE) } 'dies ok when 2 arguments passed, but 1 is incorrect in findOligo()';
-dies_ok { writeFasta(1,2,3) } 'dies ok when 3 arguments are passed in findOligo()';
+dies_ok { findOligo(1,2,3) } 'dies ok when 3 arguments are passed in findOligo()';
lives_ok { findOligo($seqDetails, $WINDOWSIZE) } 'lives ok when right paramaterers passed in findOligo()';
my ($gRNAs, $CRPseqs) = findOligo($seqDetails, $WINDOWSIZE);
@@ -89,7 +86,7 @@ is($gRNAs->{'gRNA_14'}->{'sequence'}, 'CTCCGGGCCTGCTAGATCGATGG', 'gRNA_14 sequen
is($gRNAs->{'gRNA_15'}->{'sequence'}, 'TCCGGGCCTGCTAGATCGATGGG', 'gRNA_15 sequence is correct');
is($gRNAs->{'gRNA_16'}->{'sequence'}, 'ATAGTACGTGATCACAGTCATGG', 'gRNA_16 sequence is correct');
-
+#-------------------------------------------------------------------------------
# writeFasta
print "\nTesting writeFasta()...\n";
dies_ok { writeFasta() } 'dies ok when no argument passed in writeFasta()';
@@ -98,21 +95,29 @@ dies_ok { writeFasta(1,2) } 'dies ok when 2 incorrect arguments passed in writeF
dies_ok { writeFasta(1, $OUTFILE) } 'dies ok when 2 arguments passed, but 1 is incorrect in writeFasta()';
dies_ok { writeFasta(1,2,3) } 'dies ok when 3 arguments are passed in writeFasta()';
lives_ok { writeFasta($gRNAs, $OUTFILE) } 'lives ok when right parameters are passed in writeFasta()';
-my $CRPfile = writeFasta($gRNAs, $OUTFILE);
+my $fastaFile = writeFasta($gRNAs, $OUTFILE);
+#-------------------------------------------------------------------------------
+# ss
+mkDir("$OUTDIR/ss");
+dies_ok { ss() } 'dies ok when no argument passed in ss()';
+dies_ok { ss(1) } 'dies ok when 1 argument passed in ss()';
+dies_ok { ss(1,2) } 'dies ok when 2 invalid arguments passed in ss()';
+lives_ok { ss('gRNA_0', $gRNAs->{'gRNA_0'}->{'sequence'} ) } 'lives ok when correct arguments passed in ss()';
+#-------------------------------------------------------------------------------
# Search::blast
print "\nTesting Search::blast()...\n";
dies_ok { Search::blast() } 'dies ok when no argument passed in Search::blast()';
dies_ok { Search::blast(1) } 'dies ok when 1 argument is passed in Search::blast()';
dies_ok { Search::blast(1,2,) } 'dies ok when 2 arguments passed, but at least 1 is incorrect in Search::blast()';
dies_ok { Search::blast(1,2, $OUTFILE) } 'dies ok when 3 arguments passed, but 1 is incorrect in Search::blast()';
-dies_ok { Search::blast(1, $CRPfile, $OUTFILE) } 'dies ok when 3 arguments passed, but 2 are incorrect in Search::blast()';
+dies_ok { Search::blast(1, $fastaFile, $OUTFILE) } 'dies ok when 3 arguments passed, but 2 are incorrect in Search::blast()';
dies_ok { Search::blast(1,2,3) } 'dies ok when 3 incorrect arguments are passed in Search::blast()';
-# lives_ok { Search::blast($CRPfile, \@SUBJSEQS, $OUTFILE) } 'lives ok when right parameters are passed in Search::blast()';
-# my $targets = Search::blast($CRPfile, \@SUBJSEQS, $OUTFILE);
-
+lives_ok { Search::blast($fastaFile, \@SUBJSEQS, $OUTFILE) } 'lives ok when right parameters are passed in Search::blast()';
+my $targets = Search::blast($fastaFile, \@SUBJSEQS, $OUTFILE);
+#-------------------------------------------------------------------------------
# _palindrome
print "\nTesting Search::_palindrome()...\n";
dies_ok { Search::_palindrome() } 'dies ok when no argument passed in Search::_palindrome()';
@@ -121,13 +126,44 @@ lives_ok { Search::_palindrome('ATGTA') } 'lives ok when right parameters are pa
is ( Search::_palindrome('ATGTA') , 'Yes', 'YES palindrome found is correct');
is ( Search::_palindrome('ATGTAA') , 'No', 'NO palindrome found is correct');
+#-------------------------------------------------------------------------------
+# writeResults
+print "\nTesting writeResults()...\n";
+dies_ok { writeResults() } 'dies ok when no argument passed in writeResults()';
+dies_ok { writeResults(1) } 'dies ok when 1 argument is passed in writeResults()';
+dies_ok { writeResults(1,2) } 'dies ok when 2 incorrect arguments passed in writeResults()';
+dies_ok { writeResults(1,2,3) } 'dies ok when 3 incorrect arguments passed in writeResults()';
+dies_ok { writeResults(1,2,3,4) } 'dies ok when 4 incorrect arguments passed in writeResults()';
+dies_ok { writeResults(1,2,3,4,5) } 'dies ok when 5 incorrect arguments passed in writeResults()';
+dies_ok { writeResults(1,2,3,4,5,6) } 'dies ok when 6 incorrect arguments passed in writeResults()';
+
+my $DOWNSEQ;
+my $UPSEQ;
+lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok whith both undef DOWN/UP-SEQ parameters are passed in writeResults()';
-# writeCRPfile
-# print "\nTesting writeCRPfile()...\n";
-# writeCRPfile($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE);
+$DOWNSEQ = "DWDWDW";
+lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok whith undef UPSEQ parameter is passed in writeResults()';
+$DOWNSEQ = "";
+$UPSEQ = "UPUPUP";
+lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok whith undef DOWNSEQ & defined $UPSEQ parameter is passed in writeResults()';
+$DOWNSEQ = "DWDWDW";
+$UPSEQ = "UPUPUP";
+lives_ok { writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE) } 'lives ok when right parameters are passed in writeResults()';
+writeResults($gRNAs, $targets, $DOWNSEQ, $UPSEQ, $WINDOWSIZE, $OUTFILE);
+
+#-------------------------------------------------------------------------------
+# sortIdentities
+print "\nTesting sortIdentities()...\n";
+dies_ok { sortIdentities() } 'dies ok when no arguments passed';
+dies_ok { sortIdentities(1) } 'dies ok when invalid argument passed';
+dies_ok { sortIdentities(1,2) } 'dies ok when 2 arguments passed';
+lives_ok { sortIdentities( $targets->{'gRNA_0'}{'test'}{'hsps'} ) } 'lives ok when 1 correct arguments passed';
+
+
+#-------------------------------------------------------------------------------
# Clean Up
`rm -r ./$OUTDIR`;