Mercurial > repos > big-tiandm > mirplant2
diff html_siRNA.pl @ 50:7b5a48b972e9 draft
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author | big-tiandm |
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date | Fri, 05 Dec 2014 00:11:02 -0500 |
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--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/html_siRNA.pl Fri Dec 05 00:11:02 2014 -0500 @@ -0,0 +1,788 @@ +#!/usr/bin/perl -w +#Filename: +#Author: Tian Dongmei +#Email: tiandm@big.ac.cn +#Date: 2014-5-29 +#Modified: +#Description: +my $version=1.00; + +use strict; +use Getopt::Long; +use File::Basename; + +my %opts; +GetOptions(\%opts,"i=s","format=s","o=s","h"); +if (!(defined $opts{o} and defined $opts{format} and defined $opts{i} ) || defined $opts{h}) { #necessary arguments +&usage; +} +my ($config,$prepath,$rfampath,$genomepath,$clusterpath,$annotatepath,$plotpath,$degpath); +my ($predir,$rfamdir,$genomedir,$clusterdir,$annotatedir,$plotdir,$degdir); +open IN,"<$opts{i}"; +$config=<IN>; chomp $config; +$prepath=<IN>; chomp $prepath; +$rfampath=<IN>;chomp $rfampath; +$genomepath=<IN>; chomp $genomepath; +$clusterpath=<IN>; chomp $clusterpath; +$annotatepath=<IN>; chomp $annotatepath; +$plotpath=<IN>; chomp $plotpath; +my $deg_tag=1; +if(eof){$deg_tag=0;} +else{ + $degpath=<IN>; chomp $degpath; +} +close IN; +my @tmp=split/\//,$prepath; +$predir=$tmp[-1]; +@tmp=split/\//,$rfampath; +$rfamdir=$tmp[-1]; +@tmp=split/\//,$genomepath; +$genomedir=$tmp[-1]; +@tmp=split/\//,$clusterpath; +$clusterdir=$tmp[-1]; +@tmp=split/\//,$annotatepath; +$annotatedir=$tmp[-1]; +@tmp=split/\//,$plotpath; +$plotdir=$tmp[-1]; + +my $dir=dirname($opts{'o'}); + +open OUT ,">$opts{'o'}"; +print OUT "<HTML>\n <HEAD>\n <TITLE> Analysis Report </TITLE>\n </HEAD> + <BODY bgcolor=\"lightgray\">\n <h1 align=\"center\">\n <font face=\"ºÚÌå\">\n <b>Small RNA Analysis Report</b>\n </font>\n </h1> + <h2>1. Sequence No. and quality</h2> + <h3>1.1 Sequece No.</h3> +"; + +### raw data no +open IN,"<$config"; +my @files;my @marks; my @rawNo; +while (my $aline=<IN>) { + chomp $aline; + my @tmp=split/\t/,$aline; + push @files,$tmp[0]; + + my $no=`less $tmp[0] |wc -l `; + chomp $no; + if ($opts{'format'} eq "fq" || $opts{'format'} eq "fastq") { + $no=$no/4; + } + else{ + $no=$no/2; + } + push @rawNo,$no; + + push @marks,$tmp[1]; +} +close IN; + +### preprocess +unless ($prepath=~/\/$/) { + $prepath .="/"; +} + +my @trimNo;my @collapse; +my $collapsefile=$prepath."collapse_reads.fa"; +open IN,"<$collapsefile"; +while (my $aline=<IN>) { + chomp $aline; + <IN>; + $aline=~/:([\d|_]+)_x(\d+)$/; + my @lng=split/_/,$1; + for (my $i=0;$i<@lng;$i++) { + if ($lng[$i]>0) { + $trimNo[$i] +=$lng[$i]; + $collapse[$i] ++; + } + } +} +close IN; + +my @cleanR;my @cleanT; +my $clean=$prepath."collapse_reads_18-40.fa"; +open IN,"<$clean"; +while (my $aline=<IN>) { + chomp $aline; + <IN>; + $aline=~/:([\d|_]+)_x(\d+)$/; + my @lng=split/_/,$1; + for (my $i=0;$i<@lng;$i++) { + if ($lng[$i]>0) { + $cleanR[$i] +=$lng[$i]; + $cleanT[$i] ++; + } + } +} +close IN; + +my @filterR;my @filterT; +my $filter=$prepath."collapse_reads_out.fa"; +open IN,"<$filter"; +while (my $aline=<IN>) { + chomp $aline; + <IN>; + $aline=~/:([\d|_]+)_x(\d+)$/; + my @lng=split/_/,$1; + for (my $i=0;$i<@lng;$i++) { + if ($lng[$i]>0) { + $filterR[$i] +=$lng[$i]; + $filterT[$i] ++; + } + } +} +close IN; + + +print OUT "<table border=\"1\"> +<tr align=\"center\"> +<th> </th> +"; +foreach (@marks) { + print OUT "<th> $_ </th>\n"; +} +print OUT "</tr> +<tr align=\"center\"> +<th align=\"left\">Raw Reads No. </th> +"; +foreach (@rawNo) { + print OUT "<td> $_ </td>\n"; +} +print OUT "</tr> +<tr align=\"center\"> +<th align=\"left\">Reads No. After Trimed 3\' adapter </th> +"; +foreach (@trimNo) { + print OUT "<td> $_ </td>\n"; +} +print OUT "</tr> +<tr align=\"center\"> +<th align=\"left\">Unique Tags No. </th> +"; +foreach (@collapse) { + print OUT "<td> $_ </td>\n"; +} +print OUT "</tr> +<tr align=\"center\"> +<th align=\"left\">Clean Reads No. </th> +"; +foreach (@cleanR) { + print OUT "<td> $_ </td>\n"; +} +print OUT "</tr> +<tr align=\"center\"> +<th align=\"left\">Clean Tags No. </th> +"; +foreach (@cleanT) { + print OUT "<td> $_ </td>\n"; +} +print OUT "</tr> +<tr align=\"center\"> +<th align=\"left\">Filter Reads No. \(reads count \>3\) </th> +"; +foreach (@filterR) { + print OUT "<td> $_ </td>\n"; +} +print OUT "</tr> +<tr align=\"center\"> +<th align=\"left\">Filter Tags No. \(reads count \>3\) </th> +"; +foreach (@filterT) { + print OUT "<td> $_ </td>\n"; +} +print OUT "</tr>\n</table>"; +print OUT "<p> +Note:<br /> +The raw data file path is: <b>$files[0]</b><br /> +"; +for (my $i=1;$i<@files;$i++) { + print OUT "          <b>$files[$i]</b><br />"; +} +print OUT "The collapsed file path is: <b>$collapsefile</b><br /> +The clean data file path is: <b>$clean</b><br /> +The filter (remain total reads>3) data file path is: <b>$filter</b><br /> +</p> +<h2> 1. Sequence length count</h2> +"; +print OUT "\n"; + +my $length=$prepath."length.html"; +open IN,"<$length"; +while (my $aline=<IN>) { + chomp $aline; + print OUT "$aline\n"; +} +close IN; + +print OUT "<p> Note:<br />The sequence length data: <a href=\"./$predir/reads_length_distribution_after_count_filter.txt\"> length file</a> +</p> +"; + +#### rfam +unless ($rfampath=~/\/$/) { + $rfampath .="/"; +} +unless ($genomepath=~/\/$/) { + $genomepath .="/"; +} +print OUT "<h2>2. Rfam non-miRNA annotation</h2> +<h3>2.1 Reads count</h3> +<table border=\"1\"> +<tr align=\"center\"> +"; + +my @rfamR; my @rfamT; +my $tag=1; +open IN,"<$dir/rfam_match/rfam_non-miRNA_annotation.txt"; +while (my $aline=<IN>) { + chomp $aline; + $tag=0 if($aline=~/tags\s+number/); + next if($aline=~/^\#/); + next if($aline=~/^\s*$/); + my @tmp=split/\s+/,$aline; + if($tag == 1){push @rfamR,[@tmp];} + else{push @rfamT,[@tmp];} +} +close IN; + + +print OUT "<th>RNA Name</th>\n"; +foreach (@marks) { + print OUT "<th> $_ </th>\n"; +} +for (my $i=0;$i<@rfamR;$i++) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$rfamR[$i][0]</th> + "; + for (my $j=1;$j<@{$rfamR[$i]} ;$j++) { + print OUT "<td> $rfamR[$i][$j]</td>\n"; + } +} + +print OUT "</tr>\n</table> + <h3>2.2 Tags count</h3> + <table border=\"1\"> + <tr align=\"center\"> + <th>RNA Name</th>\n"; +foreach (@marks) { + print OUT "<th> $_ </th>\n"; +} +for (my $i=0;$i<@rfamT;$i++) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$rfamT[$i][0]</th> + "; + for (my $j=1;$j<@{$rfamT[$i]} ;$j++) { + print OUT "<td> $rfamT[$i][$j]</td>\n"; + } +} +print OUT "</tr>\n</table> +<p>Note:<br />The rfam mapping results is: <b>$rfampath</b>"; +print OUT "<b>rfam_mapped.bwt</b></p>"; + +open IN,"<$dir/genome_match/genome_mapped.bwt"; +my @genome_r_u; +my @genome_r_m; +my @genome_t_u; +my @genome_t_m; +my $tags_map_number=0; +while (my $aline=<IN>) { + chomp $aline; + my @temp=split/\t/,$aline; + if ($temp[6]==0) { + $aline=~/:([\d|_]+)_x(\d+)/; + my @lng=split/_/,$1; + for (my $i=0;$i<@lng;$i++) { + if ($lng[$i]>0) { + $genome_r_u[$i] +=$lng[$i]; + $genome_t_u[$i] ++; + } + } + $tags_map_number++; + } + if ($temp[6]>0) { + $aline=~/:([\d|_]+)_x(\d+)/; + my @lng=split/_/,$1; + for (my $i=0;$i<@lng;$i++) { + if ($lng[$i]>0) { + $genome_r_m[$i] +=$lng[$i]; + $genome_t_m[$i] ++; + } + } + for (my $i=0;$i<$temp[6] ;$i++) { + my $next=<IN>; + } + $tags_map_number++; + } +} +close IN; +#<h3>3.1 Reads count</h3> +#<table border=\"1\"> +#<tr align=\"center\"> +print OUT "<h2>3. genome mapping result</h2> +<table border=\"1\"> +<tr align=\"center\"> +<th align=\"left\">Map</th>\n +"; +foreach (@marks) { + print OUT "<th> $_ </th>\n"; +} +print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Uniq Map Reads No.</th> +"; +for (my $i=0;$i<@genome_r_u ;$i++) { + print OUT "<td> $genome_r_u[$i]</td>\n"; +} + +print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Uniq Map Tags No.</th> +"; +for (my $i=0;$i<@genome_t_u ;$i++) { + print OUT "<td> $genome_t_u[$i]</td>\n"; +} + +print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Multiple Map Reads No.</th> +"; +for (my $i=0;$i<@genome_r_m ;$i++) { + print OUT "<td> $genome_r_m[$i]</td>\n"; +} + +print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Multiple Map Tags No.</th> +"; +for (my $i=0;$i<@genome_t_m ;$i++) { + print OUT "<td> $genome_t_m[$i]</td>\n"; +} + +print OUT "</tr>\n</table> +<p>Note:<br />The genome mapping results is: <b>$genomepath</b>"; +print OUT "<b>genome_mapped.bwt</b></p>"; + +my $cluster="$clusterpath/sample_reads.cluster"; +my $cluster_number=`less $cluster |wc -l `; +$cluster_number=$cluster_number-1; +my (%cluster_length,@exp,@rpkm); +my @exp_range=qw(0 \(0--10] \(10--100] \(100--1000] \(1000--10000] \(10000--100000] \(100000--**\)); +my @rpkm_range=qw(0 \(0--0.25] \(0.25--0.5] \(0.5--1] \(1.0-5.0] \(5--10] \(10--50] \(50--100] \(100--500] \(500--1000] \(1000--**]); + +open IN,"<$cluster"; +while (my $aline=<IN>) { + next if($aline=~/^\"/); + chomp $aline; + my @temp=split/\t/,$aline; + my @id=split/:|-/,$temp[0]; + $cluster_length{$id[2]-$id[1]+1}++; + for (my $i=0;$i<@marks ;$i++) { + if ($temp[$i+3] == 0) {$exp[0][$i]++;} + elsif ($temp[$i+3]>0 && $temp[$i+3]<= 10 ){$exp[1][$i]++;} + elsif ($temp[$i+3]>10 && $temp[$i+3]<=100){$exp[2][$i]++;} + elsif ($temp[$i+3]>100 && $temp[$i+3]<=1000){$exp[3][$i]++;} + elsif ($temp[$i+3]>1000 && $temp[$i+3]<=10000){$exp[4][$i]++;} + elsif ($temp[$i+3]>10000 && $temp[$i+3]<=100000){$exp[5][$i]++;} + elsif ($temp[$i+3]>100000){$exp[6][$i]++;} + } +} +close IN; + +my $cluster_rpkm="$clusterpath/sample_rpkm.cluster"; +open IN,"<$cluster_rpkm"; +while (my $aline=<IN>) { + next if($aline=~/^\#/); + chomp $aline; + my @temp=split/\t/,$aline; + for (my $i=0;$i<@marks ;$i++) { + if ($temp[$i+3]==0) {$rpkm[0][$i]++;} + elsif($temp[$i+3]>0 && $temp[$i+3]<=0.25){$rpkm[1][$i]++;} + elsif($temp[$i+3]>0.25 && $temp[$i+3]<=0.5){$rpkm[2][$i]++;} + elsif($temp[$i+3]>0.5 && $temp[$i+3]<=1){$rpkm[3][$i]++;} + elsif($temp[$i+3]>1 && $temp[$i+3]<=5){$rpkm[4][$i]++;} + elsif($temp[$i+3]>5 && $temp[$i+3]<=10){$rpkm[5][$i]++;} + elsif($temp[$i+3]>10 && $temp[$i+3]<=50){$rpkm[6][$i]++;} + elsif($temp[$i+3]>50 && $temp[$i+3]<=100){$rpkm[7][$i]++;} + elsif($temp[$i+3]>100 && $temp[$i+3]<=500){$rpkm[8][$i]++;} + elsif($temp[$i+3]>500 && $temp[$i+3]<=1000){$rpkm[9][$i]++;} + else{$rpkm[10][$i]++;} + } +} + +close IN; + +my $cluster_length_file="$clusterpath/cluster_length.txt"; +open LEN,">$cluster_length_file"; +print LEN "\#length\tcluster_number\n"; +foreach my $key (sort keys %cluster_length) { + print LEN "$key\t$cluster_length{$key}\n"; +} +close LEN; +print OUT "<h2>4. cluster result</h2> +<h3>4.1 Cluster count</h3> +<table border=\"1\"> +<tr align=\"center\"> +<th align=\"left\"> </th> +<td>Merged samples</td></tr> +<tr align=\"center\"> +<th align=\"left\">Tags number</th> +<td>$tags_map_number</td></tr> +<tr align=\"center\"> +<th align=\"left\">Cluster number</th> +<td>$cluster_number</td></tr>\n</table> +"; + +print OUT "<h3>4.2 Cluster length</h3> +<p> Note:<br />The clusters length data: <a href=\"./$clusterdir/cluster_length.txt\"> length file</a> +</p> +"; +print OUT "<h3>4.3 Quantify</h3> +<table border=\"1\"> +<tr align=\"center\"> +<th align=\"left\">Reads Range</th>\n +"; +foreach (@marks) { + print OUT "<th> $_ </th>\n"; +} +for (my $i=0;$i<@exp_range;$i++) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$exp_range[$i]</th> + "; + for (my $j=0;$j<@marks ;$j++) { + if (!(defined($exp[$i][$j]))) { + print OUT "<td> 0</td>\n"; + } + else{print OUT "<td> $exp[$i][$j]</td>\n";} + } +} +print OUT "</tr>\n</table>"; + +print OUT "\n<table border=\"1\"> +<tr align=\"center\"> +<th align=\"left\">RPKM Range</th>\n +"; +foreach (@marks) { + print OUT "<th> $_ </th>\n"; +} +for (my $i=0;$i<@rpkm_range;$i++) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$rpkm_range[$i]</th> + "; + for (my $j=0;$j<@marks ;$j++) { + if (!(defined($rpkm[$i][$j]))) { + print OUT "<td> 0</td>\n"; + } + else{print OUT "<td> $rpkm[$i][$j]</td>\n";} + } +} +print OUT "</tr>\n</table>"; + +my $annotate="$annotatepath/sample_c_p.anno"; +my (%posit,%repeat,%nat1,%nat2); +my (@phase,@long,@repeat,@nat); +for (my $j=0;$j<@marks ;$j++) { + $phase[$j]=0; + $long[$j]=0; + $repeat[$j]=0; + $nat[$j]=0; +} + +my $class_anno=1; +open ANNO,"<$annotate"; +while (my $aline=<ANNO>) { + chomp $aline; + my @temp=split/\t/,$aline; + if($aline=~/^\#/){ + if (@temp != 10+@marks) { + $class_anno=0; + } + next; + } + for (my $i=3+@marks+$class_anno;$i<@temp;$i++) { + my @posit=split/\;/,$temp[$i]; + for (my $j=0;$j<@marks ;$j++) { + if ($temp[3+$j]>0) { + $posit{$posit[0]}[$j]++; + } + else{ + if (!(defined($posit{$posit[0]}[$j]))) { + $posit{$posit[0]}[$j]=0; + } + } + } + } + if ($class_anno) { + for (my $j=0;$j<@marks ;$j++) { + if ($temp[3+$j]>0) { + if ($temp[6] eq "phase") { + $phase[$j]++; + } + if ($temp[7] eq "long") { + $long[$j]++; + } + if ($temp[8] ne "\/") { + $repeat[$j]++; + my @rr=split/\;/,$temp[8]; + foreach (@rr) { + $repeat{$_}[$j]++; + } + } + if ($temp[9] ne "\/") { + $nat[$j]++; + my @nn1=split/\;/,$temp[9]; + my @nn2=split/\;/,$temp[10]; + for (my $k=0;$k<@nn1 ;$k++) { + $nat1{$nn1[$k]}[$j]++; + $nat2{$nn2[$k]}[$j]++; + } + } + } + } + } +} +close ANNO; + +print OUT "<h2>5. Cluster Annotate</h2> +<h3>5.1 Cluster genome position annotate</h3> +<table border=\"1\"> +<tr align=\"center\"> +<th align=\"left\">clusters number</th>\n +"; + +foreach (@marks) { + print OUT "<th> $_ </th>\n"; +} +foreach my $key (sort keys %posit) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$key</th> + "; + foreach (@{$posit{$key}}) { + print OUT "<td> $_</td>\n"; + } +} +print OUT "</tr>\n</table>"; +print OUT "<p> +Note:<br /> +One cluster mybe annotate to multiple genes<br /> +"; + +if ($class_anno) { + print OUT "<h3>5.2 Cluster source mechanism annotate</h3> + <table border=\"1\"> + <tr align=\"center\"> + <th align=\"left\">clusters number</th>\n + "; + + foreach (@marks) { + print OUT "<th> $_ </th>\n"; + } + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Phase</th>\n + "; + foreach (@phase) { + print OUT "<td> $_ </td>\n"; + } + + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Long</th>\n + "; + foreach (@long) { + print OUT "<td> $_ </td>\n"; + } + + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Repeat</th>\n + "; + foreach (@repeat) { + print OUT "<td> $_ </td>\n"; + } + + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">Nat</th>\n + "; + foreach (@nat) { + print OUT "<td> $_ </td>\n"; + } + print OUT "</tr>\n</table>"; + + print OUT "<p> + Repeat subclass annotate: + "; + + print OUT "<table border=\"1\"> + <tr align=\"center\"> + <th align=\"left\">Repeat subclass</th>\n + "; + foreach (@marks) { + print OUT "<th> $_ </th>\n"; + } + + foreach my $key (sort keys %repeat) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$key</th> + "; + for (my $i=0;$i<@marks ;$i++) { + if (defined($repeat{$key}[$i])) { + print OUT "<td> $repeat{$key}[$i] </td>\n"; + } + else{print OUT "<td> 0 </td>\n";} + } + } + print OUT "</tr>\n</table>"; + + + print OUT "<p> + Nat subclass1 annotate: + "; + + print OUT "<table border=\"1\"> + <tr align=\"center\"> + <th align=\"left\">Nat subclass1</th>\n + "; + foreach (@marks) { + print OUT "<th> $_ </th>\n"; + } + foreach my $key (sort keys %nat1) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$key</th> + "; + for (my $i=0;$i<@marks ;$i++) { + if (defined($nat1{$key}[$i])) { + print OUT "<td> $nat1{$key}[$i] </td>\n"; + } + else{print OUT "<td> 0 </td>\n";} + } + } + print OUT "</tr>\n</table>"; + + print OUT "<p> + Nat subclass2 annotate: + "; + + print OUT "<table border=\"1\"> + <tr align=\"center\"> + <th align=\"left\">Nat subclass2</th>\n + "; + foreach (@marks) { + print OUT "<th> $_ </th>\n"; + } + foreach my $key (sort keys %nat2) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$key</th> + "; + for (my $i=0;$i<@marks ;$i++) { + if (defined($nat2{$key}[$i])) { + print OUT "<td> $nat2{$key}[$i] </td>\n"; + } + else{print OUT "<td> 0 </td>\n";} + } + } + print OUT "</tr>\n</table>"; + print OUT "<p> + Note:<br /> + One cluster mybe annotate to multiple repeats or nats<br /> + "; +} +else { + print OUT "<h3>5.2 Cluster source mechanism annotate</h3> + <br />Do not do source mechanism annotate <br />"; + +} + +print OUT "<h2>6. Graph of Clusters of all samples</h2> \n"; + +my $plot=$plotpath."cluster.html"; +open IN,"<$plot"; +while (my $aline=<IN>) { + chomp $aline; + print OUT "$aline\n"; +} +close IN; + + +if ($deg_tag) { + my $deg_file=`ls $degpath`; + chomp $deg_file; + my @deg_file=split/\n/,$deg_file; + my %deg; + foreach (@deg_file) { + my $output="$degpath/$_/output_score.txt"; + open IN,"<$output"; + $deg{$_}[0]=0; + $deg{$_}[1]=0; + $deg{$_}[2]=0; + while (my $aline=<IN>) { + next if ($aline=~/^\"/); + chomp $aline; + my @temp=split/\t/,$aline; + if ($temp[9] eq "TRUE") { + $deg{$_}[0]++; + if ($temp[4] >0) { + $deg{$_}[1]++; + } + if ($temp[4] <0) { + $deg{$_}[2]++; + } + } + } + close IN; + } + + print OUT "<h2>7. DEG</h2> + <table border=\"1\"> + <tr align=\"center\"> + <th align=\"left\">Genes number</th>\n + <th> DEG </th>\n + <th> UP </th>\n + <th> DOWN </th>\n + "; + + foreach my $key (sort keys %deg) { + print OUT "</tr> + <tr align=\"center\"> + <th align=\"left\">$key</th> + "; + for (my $i=0;$i<@{$deg{$key}} ;$i++) { + print OUT "<td> $deg{$key}[$i] </td>\n"; + } + } + print OUT "</tr>\n</table>"; +} +else{ + print OUT "<h2>7. DEG</h2> + <br />Do not do DE clusters <br />"; +} + +print OUT " + </BODY> +</HTML> +"; +close OUT; + + + + +sub usage{ +print <<"USAGE"; +Version $version +Usage: +$0 -o +options: +-i +-format +-o output file +-h help +USAGE +exit(1); +}