#@UGENE_WORKFLOW #Call variants for an input assembly and a reference sequence using SAMtools mpileup and bcftool. #Predict effects of the found variants using SnpEff. workflow "Variant calling and effects prediction"{ read-assembly { type:get-file-list; name:"Read Assembly (BAM/SAM)"; url-in { dataset:Dataset; } } read-sequence { type:get-file-list; name:"Read Sequence"; url-in { dataset:Dataset; } } call_variants { type:call_variants; name:"Call Variants"; max-dep:100; reference-source:port; variants-url:variants.vcf; } files-conversion-1 { type:files-conversion; name:"To FASTA"; document-format:fasta; } files-conversion-2 { type:files-conversion; name:"To BAM"; document-format:bam; } rename-chromosome-in-variation { type:rename-chromosome-in-variation; name:"Chr Notation"; prefix-replace-with:chr; prefixes-to-replace:"NC_00000;NC_0000"; } seff { type:seff; name:"Annotate and Predict Effects with SnpEff"; } convert-snpeff-variations-to-annotations { type:convert-snpeff-variations-to-annotations; name:"To Annotations"; } .actor-bindings { call_variants.out-variations->rename-chromosome-in-variation.in-file files-conversion-2.out-file->call_variants.in-assembly read-sequence.out-url->files-conversion-1.in-file files-conversion-1.out-file->call_variants.in-sequence seff.out-file->convert-snpeff-variations-to-annotations.in-variations-url read-assembly.out-url->files-conversion-2.in-file rename-chromosome-in-variation.out-file->seff.in-file } read-assembly.dataset->call_variants.in-assembly.dataset files-conversion-2.url->call_variants.in-assembly.url files-conversion-1.url->call_variants.in-sequence.url read-sequence.url->files-conversion-1.in-file.url read-assembly.url->files-conversion-2.in-file.url call_variants.url->rename-chromosome-in-variation.in-file.url rename-chromosome-in-variation.url->seff.in-file.url seff.url->convert-snpeff-variations-to-annotations.in-variations-url.url .meta { parameter-aliases { read-assembly.url-in { alias:assembly_in; } read-sequence.url-in { alias:seq_in; } call_variants.acgt_only { alias:a_c_g_t; } call_variants.bcf_bed { alias:bcf_bed; } call_variants.bed { alias:bed_pos_list_file; } call_variants.disable_baq { alias:BAQ; } call_variants.keepalt { alias:retain_all_pos_alt; } call_variants.max_depth { alias:max_reads_num; } call_variants.no_indel { alias:no_indels; } call_variants.ptype { alias:allele_freq_spect; } call_variants.theta { alias:theta; } call_variants.variants-url { alias:out_file; } } visual { call_variants { pos:"-465 -510"; style:ext; bg-color-ext:"0 129 94 64"; bounds:"-30 -30 123.125 72"; in-assembly.angle:176.73; in-sequence.angle:235.098; out-variations.angle:351.674; } convert-snpeff-variations-to-annotations { pos:"-450 -285"; style:simple; bg-color-simple:"234 223 240 255"; in-variations-url.angle:3.25195; } files-conversion-1 { pos:"-615 -450"; style:simple; bg-color-simple:"84 84 84 255"; in-file.angle:196.858; out-file.angle:6.5198; } files-conversion-2 { pos:"-615 -555"; style:simple; bg-color-simple:"84 84 84 255"; in-file.angle:170.538; out-file.angle:334.799; } read-assembly { pos:"-870 -600"; style:ext; bg-color-ext:"24 102 175 64"; bounds:"-30 -30 111.375 66"; out-url.angle:360; } read-sequence { pos:"-885 -435"; style:ext; bg-color-ext:"24 102 175 64"; bounds:"-30 -30 110.25 62"; out-url.angle:349.061; } rename-chromosome-in-variation { pos:"-210 -495"; style:simple; bg-color-simple:"232 227 230 255"; in-file.angle:180; out-file.angle:270.785; } seff { pos:"-270 -285"; style:ext; bg-color-ext:"0 129 94 64"; in-file.angle:28.6105; out-file.angle:181.848; } call_variants.out-variations->rename-chromosome-in-variation.in-file { text-pos:"-45 -43"; } files-conversion-1.out-file->call_variants.in-sequence { text-pos:"-19 -31"; } files-conversion-2.out-file->call_variants.in-assembly { text-pos:"-12 -24"; } read-assembly.out-url->files-conversion-2.in-file { text-pos:"-37 -11"; } read-sequence.out-url->files-conversion-1.in-file { text-pos:"-31 -21"; } rename-chromosome-in-variation.out-file->seff.in-file { text-pos:"-45 -43"; } seff.out-file->convert-snpeff-variations-to-annotations.in-variations-url { text-pos:"-45 -43"; } } wizard { auto-run: true; name:"Call Variants Wizard"; page { id:1; next:2; title:"Input reference sequence and assembly"; parameters-area { group { title:"Input files"; lineedit-and-dataset { read-sequence.url-in { label:"Reference sequence file"; } read-assembly.url-in { label:"BAM/SAM file"; } } } } } page { id:2; next:3; title:"SAMtools mpileup parameters"; parameters-area { group { title:Parameters; label-size:200; call_variants.use_orphan { } call_variants.disable_baq { } call_variants.capq_thres { } call_variants.max_depth { } call_variants.ext_baq { } call_variants.bed { } call_variants.reg { } call_variants.min_mq { } call_variants.min_baseq { } } group { title:Additional; label-size:200; type:hideable; call_variants.illumina13-encoding { } call_variants.extQ { } call_variants.tandemQ { } call_variants.no_indel { } call_variants.max_indel_depth { } call_variants.openQ { } call_variants.pl_list { } } } } page { id:3; next:4; title:"SAMtools bcftools view parameters"; parameters-area { group { title:Parameters; label-size:200; call_variants.keepalt { } call_variants.fix_pl { } call_variants.no_geno { } call_variants.acgt_only { } call_variants.bcf_bed { } call_variants.qcall { } call_variants.samples { } call_variants.min_smpl_frac { } call_variants.call_gt { } } group { title:Additional; label-size:200; type:hideable; call_variants.indel_frac { } call_variants.pref { } call_variants.ptype { } call_variants.theta { } call_variants.ccall { } call_variants.n1 { } call_variants.n_perm { } call_variants.min_perm_p { } } } } page { id:4; next:5; title:"SAMtools vcfutils varFilter parameters"; parameters-area { group { title:Parameters; label-size:200; call_variants.print-filtered { } call_variants.min-qual { } call_variants.min-dep { } call_variants.max-dep { } call_variants.min-alt-bases { } call_variants.gap-size { } call_variants.window { } } group { title:Additional; label-size:200; type:hideable; call_variants.min-strand { } call_variants.min-baseQ { } call_variants.min-mapQ { } call_variants.min-end-distance { } call_variants.min-hwe { } } } } page { id:5; next:6; title:"Change chromosome notation for variations"; parameters-area { group { title:Parameters; label-size:150; rename-chromosome-in-variation.prefixes-to-replace { } rename-chromosome-in-variation.prefix-replace-with { } } } } page { id:6; next:7; title:"SnpEff parameters"; parameters-area { group { title:Parameters; label-size:200; seff.genome { } } group { title:Additional; label-size:200; type:hideable; seff.canon { } seff.hgvs { } seff.lof { } seff.motif { } seff.updown-length { } } } } page { id:7; title:"Output files"; parameters-area { group { title:"Output variants"; label-size:110; call_variants.variants-url { label:"Output VCF file" } convert-snpeff-variations-to-annotations.url-out { label:"Output GenBank file" } } } } } } }