Merge pull request #3 from fmalmeida/dev

added minimap and options to configure plot
fmalmeida · Dec 20, 2022 · bcd581f · bcd581f
2 parents e1c52c2 + c8a4ef9
commit bcd581f
Show file tree

Hide file tree

Showing 10 changed files with 118 additions and 19 deletions.
diff --git a/recipe/bin/plot_circos b/recipe/bin/plot_circos
@@ -1,4 +1,4 @@
-#!/bin/bash
+#!/usr/bin/env bash
 # Simple script to create a circos plot between two FASTA files.
 
 ######################
@@ -34,6 +34,12 @@ export GCWINDOW=5000
 export GCSTEP=5000
 export BACANNOT="no"
 export SKIP_LINKS="no"
+export USE_MINIMAP2="no"
+export MINIMAP2_METHOD="asm20"
+export MAX_TICKS="5000"
+export MAX_IDEOGRAMS="200"
+export MAX_LINKS="50000"
+export MAX_POINTS_PER_TRACK="50000"
 
 ######################################
 ### Function to filter FASTA files ###
@@ -55,6 +61,11 @@ source ${SCRIPT_DIR}/../src/find_links.sh
 #########################################################################
 source ${SCRIPT_DIR}/../src/parse_links.sh
 
+########################################
+### Function to use minimap2 instead ###
+########################################
+source ${SCRIPT_DIR}/../src/minimap2.sh
+
 #############################################################################
 ### Function to sort and remove duplicates from links and karyotype files ###
 #############################################################################
@@ -234,6 +245,30 @@ case $ARGS in
       export SKIP_LINKS="yes"
       shift
       ;;
+    --use_minimap2)
+      export USE_MINIMAP2="yes"
+      shift
+      ;;
+    --minimap2_method)
+      export MINIMAP2_METHOD=$2
+      shift 2
+      ;;
+    --max_ticks)
+      export MAX_TICKS=$2
+      shift 2
+      ;;
+    --max_ideograms)
+      export MAX_IDEOGRAMS=$2
+      shift 2
+      ;;
+    --max_links)
+      export MAX_LINKS=$2
+      shift 2
+      ;;
+    --max_points_per_track)
+      export MAX_POINTS_PER_TRACK=$2
+      shift 2
+      ;;
     *)
       printf "******************************\n"
       printf "Error: Invalid argument $1\n"

diff --git a/recipe/meta.yaml b/recipe/meta.yaml
@@ -1,5 +1,5 @@
 {% set name = "easy_circos" %}
-{% set version = "0.3" %}
+{% set version = "0.4" %}
 
 package:
   name: "{{ name|lower }}"
@@ -22,6 +22,7 @@ requirements:
     - perl-app-cpanminus
     - perl-module-build
     - blast
+    - minimap2
   run:
     - perl
     - circos
@@ -32,6 +33,7 @@ requirements:
     - perl-app-cpanminus
     - perl-module-build
     - blast
+    - minimap2
 
 about:
   home: "https://github.com/fmalmeida/easy_circos_plot"

diff --git a/recipe/src/filter.sh b/recipe/src/filter.sh
@@ -13,5 +13,4 @@ while read -r FASTA FASTA_PREFIX FASTA_COLOR ; do
   $CONDA_PREFIX/bin/perl $CONDA_PREFIX/bin/removesmalls.pl $MINLEN $FASTA >> ${RESULTS}/filtered/"$name" ;
   continue
 done<"$FOFN"
-
 }
diff --git a/recipe/src/find_links.sh b/recipe/src/find_links.sh
@@ -1,22 +1,22 @@
 find_links()
 {
 # create dir
-mkdir -p ${RESULTS}/all_vs_all_blast
+mkdir -p ${RESULTS}/all_vs_all_links
 
 # concatenate genomes
 cat ${RESULTS}/filtered/* >> ${RESULTS}/concatenated_genomes.fasta ;
 export CONCAT_FASTA=${RESULTS}/concatenated_genomes.fasta
-export BLAST_DB=${RESULTS}/all_vs_all_blast/blast_db
+export BLAST_DB=${RESULTS}/all_vs_all_links/blast_db
 
 # Run blast
 makeblastdb -in $CONCAT_FASTA -dbtype nucl -out $BLAST_DB &> /dev/null ;
 blastn -task blastn -perc_identity $MINID -query $CONCAT_FASTA -db $BLAST_DB \
     -outfmt "6 qseqid qstart qend sseqid sstart send pident length mismatch gapopen evalue bitscore stitle" \
-    -out ${RESULTS}/all_vs_all_blast/tmp.blast -num_threads $THREADS
+    -out ${RESULTS}/all_vs_all_links/tmp.blast -num_threads $THREADS
 
 # Filter blast
-awk -F '\t' -v minid=$MINID '{ if ($7 >= minid) { print } }' ${RESULTS}/all_vs_all_blast/tmp.blast > ${RESULTS}/all_vs_all_blast/all_vs_all.blast
+awk -F '\t' -v minid=$MINID '{ if ($7 >= minid) { print } }' ${RESULTS}/all_vs_all_links/tmp.blast > ${RESULTS}/all_vs_all_links/all_vs_all.aln.txt
 
 # Remove tmp
-rm ${RESULTS}/all_vs_all_blast/tmp.blast
+rm ${RESULTS}/all_vs_all_links/tmp.blast
 }
diff --git a/recipe/src/help.sh b/recipe/src/help.sh
@@ -33,13 +33,20 @@ Copyright, Felipe Almeida <almeidafmarques@outlook.com>, 2021
     # Input min. length
     --minlen                            Min size of contigs to consider for plot [Default: 10000]
 
-    # Links (blastn) min. percentage id
+    # Links configurations
     --skip_links                        Do not compute blast and do not draw links.
                                         Useful for when only desiring the configs. [Default: false]
+    
+    --use_minimap2                      Compute links with minimap2. This is only useful for big genomes.
+                                        With small genomes probably no align block will be detected.
+                                        For minimap2, --minid and --linklen have no effect. By default uses blastn.
+    
+    --minimap2_method                   Select alignment "method" / "algorithm" for minimap2.
+                                        Options: asm5 | asm10 | asm20. [Default: asm20]
 
-    --minid                             Min. percentage id to filter the results of blastn to draw links [Default: 85]
+    --minid                             Min. percentage id to filter the results of blastn (only for blastn) to draw links [Default: 85]
 
-    --linklen                           Min. link (blastn hit) length to display in plot [Default: 5000]
+    --linklen                           Min. length of blastn hits (only for blastn) length to display in plot [Default: 5000]
     
     --show_intrachr                     Tells the program to create a conf file showing intra chr links [Default: false]
                                         Mandatory if using only one FASTA, otherwise, links will not be shown.
@@ -60,6 +67,13 @@ Copyright, Felipe Almeida <almeidafmarques@outlook.com>, 2021
                                         3 or 4 columns as shown at http://circos.ca/documentation/tutorials/configuration/data_files.
                                         The first column must be the name (ID) of the contig.
                                         Checkout the "--gff2tiles" script (below).
+    
+    # Housekeeping conf
+
+    --max_ticks                         Max number of ticks allowed to plot. [Default: 5000]
+    --max_ideograms                     Max number of ideograms allowed to plot. [Default: 200]
+    --max_links                         Max number of links allowed to plot. [Default: 50000]
+    --max_points_per_track              Max number of points per track (e.g. histogram) allowed to plot. [Default: 50000]
 
 
     # Helpful scripts!

diff --git a/recipe/src/minimap2.sh b/recipe/src/minimap2.sh
@@ -0,0 +1,33 @@
+minimap_links()
+{
+    # create dir
+    mkdir -p ${RESULTS}/all_vs_all_links ${RESULTS}/conf
+
+    # concatenate genomes
+    cat ${RESULTS}/filtered/* >> ${RESULTS}/concatenated_genomes.fasta ;
+    export CONCAT_FASTA=${RESULTS}/concatenated_genomes.fasta
+
+    # run minimap2
+    minimap2 \
+        -c --cs -t $THREADS \
+        -x asm20 -o ${RESULTS}/all_vs_all_links/all_vs_all.aln.txt \
+        $CONCAT_FASTA $CONCAT_FASTA 2> /dev/null
+
+    # parse paf
+    cut -f 1,3,4,6,8,9 ${RESULTS}/all_vs_all_links/all_vs_all.aln.txt >> ${RESULTS}/conf/links_concatenated.txt
+
+    # get links comming from contigs and give it colors
+    IFS=','
+    while read -r FASTA FASTA_PREFIX FASTA_COLOR ; do
+    bioawk -c fastx '{ printf $name"\n" }' $FASTA > tmp_names.fasta ;
+    awk -v color1=$FASTA_COLOR -F'\t' 'NR==FNR{c[$1]++;next};c[$1] > 0 {print $0 "\t" "color="color1}' \
+        tmp_names.fasta ${RESULTS}/conf/links_concatenated.txt >> ${RESULTS}/conf/links_concatenated_colored.txt
+    rm tmp_names.fasta ;
+    done<"$FOFN"
+
+    # create additional file whithout intrachr links
+    awk \
+        -F'\t' \
+        '{ if ($1 != $4) { print } }' \
+        ${RESULTS}/conf/links_concatenated_colored.txt > ${RESULTS}/conf/links_concatenated_colored_no_intrachr.txt ;
+}
diff --git a/recipe/src/parse_links.sh b/recipe/src/parse_links.sh
@@ -5,7 +5,7 @@ mkdir -p ${RESULTS}/conf
 
 # Filter blocks with more then N bp hits
 awk -v minlen=$MINLINKLEN '{ if ($8 >= minlen) { print } }' \
-  ${RESULTS}/all_vs_all_blast/all_vs_all.blast | cut -f 1,2,3,4,5,6 >> ${RESULTS}/conf/links_concatenated.txt ;
+  ${RESULTS}/all_vs_all_links/all_vs_all.aln.txt | cut -f 1,2,3,4,5,6 >> ${RESULTS}/conf/links_concatenated.txt ;
 
 # get links comming from contigs and give it colors
 IFS=','
@@ -25,7 +25,7 @@ empty_links()
   # concatenate genomes
   cat ${RESULTS}/filtered/* >> ${RESULTS}/concatenated_genomes.fasta ;
   export CONCAT_FASTA=${RESULTS}/concatenated_genomes.fasta
-  export BLAST_DB=${RESULTS}/all_vs_all_blast/blast_db
+  export BLAST_DB=${RESULTS}/all_vs_all_links/blast_db
 
   touch ${RESULTS}/conf/links_concatenated_colored.txt ${RESULTS}/conf/links_concatenated_colored_no_intrachr.txt 
 }
diff --git a/recipe/src/plot_circos.sh b/recipe/src/plot_circos.sh
@@ -6,6 +6,14 @@ CURRENT_DIR=$PWD
 # got to conf dir
 cd ${RESULTS}/conf/
 
+# get etc/files
+cp -r $(which circos | sed 's|/circos$|/../etc|g') .
+
+sed -i "s/max_ticks.*= 5000/max_ticks = ${MAX_TICKS}/" etc/housekeeping.conf
+sed -i "s/max_ideograms.*= 200/max_ideograms = ${MAX_IDEOGRAMS}/" etc/housekeeping.conf
+sed -i "s/max_links.*= 25000/max_links = ${MAX_LINKS}/" etc/housekeeping.conf
+sed -i "s/max_points_per_track.*= 25000/max_points_per_track = ${MAX_POINTS_PER_TRACK}/" etc/housekeeping.conf
+
 # draw
 circos
 

diff --git a/recipe/src/workflow.sh b/recipe/src/workflow.sh
@@ -14,9 +14,17 @@ workflow()
   # Step 3
   if [ "$SKIP_LINKS" = "no" ]
   then
-  echo " # Finding links (all vs all blastn)!"
-  find_links      ;
-  parse_links     ;
+
+    if [ "$USE_MINIMAP2" = "yes" ]
+    then
+    echo " # Finding links (all vs all minimap2)!"
+    minimap_links   ;
+    else
+    echo " # Finding links (all vs all blastn)!"
+    find_links      ;
+    parse_links     ;
+    fi
+
   else
   echo " # Skipping links (all vs all blastn)!"
   empty_links     ;

diff --git a/recipe/src/write_circos.sh b/recipe/src/write_circos.sh
@@ -3,6 +3,9 @@ write_circos()
 cat << EOF
 # MINIMUM CIRCOS CONFIGURATION
 
+# debugging, I/O an dother system parameters
+<<include etc/housekeeping.conf>> # included from Circos distribution
+
 # Defines unit length for ideogram and tick spacing, referenced
 # using "u" prefix, e.g. 10u
 chromosomes_units = 500000
@@ -94,9 +97,6 @@ ${TILES_CONF}
 
 </plots>
 
-# debugging, I/O an dother system parameters
-<<include etc/housekeeping.conf>> # included from Circos distribution
-
 show_ticks = yes
 show_tick_labels = yes