{"id":2192,"date":"2016-12-12T14:50:24","date_gmt":"2016-12-12T12:50:24","guid":{"rendered":"http:\/\/bioinfo2.ugr.es\/MethFlow\/?page_id=2192"},"modified":"2016-12-16T10:58:17","modified_gmt":"2016-12-16T08:58:17","slug":"quick-start","status":"publish","type":"page","link":"https:\/\/bioinfo2.ugr.es\/MethFlow\/quick-start\/","title":{"rendered":"Quick start"},"content":{"rendered":"<p><span id=\"ouHighlight__131_134TO132_134\">Quick start is thought to try MethFlow without going into details and using the MethFlow virtual machine. For further information, see<\/span><span id=\"noHighlight_0.03053827676673926\">\u00a0<\/span><span id=\"ouHighlight__162_163TO158_160\">the<\/span><span id=\"noHighlight_0.34981369552068187\">\u00a0<a href=\"http:\/\/bioinfo2.ugr.es\/MethFlow\/reference-manual\/\">reference manual<\/a><\/span><span id=\"noHighlight_0.8654089653028183\">.<\/span><\/p>\n<h3>Install MethFlow<sup>VM<\/sup><\/h3>\n<ol>\n<li>Install <a href=\"https:\/\/www.virtualbox.org\/wiki\/Downloads\">VirtualBox<\/a>.<\/li>\n<li>Install <a href=\"https:\/\/www.virtualbox.org\/wiki\/Downloads\">VirtualBox Extension Pack<\/a>.<\/li>\n<li>Download <a href=\"http:\/\/bioinfo2.ugr.es\/MethFlow\/download\">MethFlow<sup>VM<\/sup><\/a>\u00a0(<a href=\"https:\/\/docs.google.com\/uc?id=0B6zaHLTx5o2bUWhPOXN0X1F4aEU&amp;export=download\">mirror<\/a>).<\/li>\n<li>Import MethFlow<sup>VM<\/sup>\u00a0to VirtualBox by double-clicking.<\/li>\n<li>Optional: <a href=\"https:\/\/www.virtualbox.org\/manual\/ch04.html#sharedfolders\" target=\"_blank\">add a shared folder<\/a> (strongly recommended).<\/li>\n<li>Run MethFlow<sup>VM<\/sup><\/li>\n<\/ol>\n<h3>The local database<\/h3>\n<h4><strong>Set your\u00a0working\u00a0folder<\/strong><\/h4>\n<p>At first startup, you will be asked which working folder you want to use. If you ignore this question, your home folder, <em>\/home\/methflow<\/em>, will be used as working folder.<\/p>\n<p>If you want to change the working folder, open a terminal and type the following command:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow_manager working_folder<\/strong><\/pre>\n<h4><strong>Set your assembly collection<\/strong><\/h4>\n<p>Tell MethFlow where the assembly collection is by typing:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow_manager assembly_collection Assemblies<\/strong><\/pre>\n<p style=\"text-align: left;\">This command\u00a0looks for\u00a0a folder named\u00a0<em>Assemblies<\/em>\u00a0inside the working folder.<\/p>\n<h4><strong>Set your\u00a0adapter collection<\/strong><\/h4>\n<p>Tell MethFlow where the adapter collection is by typing:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow_manager adapter_collection Adapters<\/strong><\/pre>\n<p>This command\u00a0looks for\u00a0a folder named\u00a0<em>Adapters<\/em>\u00a0inside the working folder.<\/p>\n<h4><strong>Set your root input folder<\/strong><\/h4>\n<p>Tell MethFlow where to\u00a0look for\u00a0the input folders:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow_manager root_input_folder Inputs<\/strong><\/pre>\n<p style=\"text-align: left;\">This command\u00a0looks for\u00a0a folder named\u00a0<em>Inputs<\/em>\u00a0inside the working folder.<\/p>\n<h4><strong>Set your root output folder<\/strong><\/h4>\n<p>Tell MethFlow where to kept the output folders:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow_manager root_output_folder Outputs<\/strong><\/pre>\n<p style=\"text-align: left;\">This command creates a folder named\u00a0<em>Outputs<\/em> inside the working folder.<\/p>\n<h3><strong>Get test datasets<\/strong><\/h3>\n<p>Open a terminal and type the following command to get test datasets:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow_manager get_test_datasets<\/strong><\/pre>\n<p>Test datasets are then downloaded and unpacked.<\/p>\n<p>These test datasets contain:<\/p>\n<ul>\n<li>The collection of adapters of <a href=\"http:\/\/www.usadellab.org\/cms\/?page=trimmomatic\"><em>Trimmomatic<\/em><\/a>. It goes to adapter collection.<\/li>\n<li>A small assembly (chromosomes 12 and 19 of hg38). It goes to assemby collection.<\/li>\n<li>Data from nine samples (from three individuals and three tissues). It goes to root input folder.<\/li>\n<\/ul>\n<h3><strong>Launch MethFlow<\/strong><\/h3>\n<p>Now, launch MethFlow with default options:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow<\/strong><\/pre>\n<p>This command looks inside the working folder and asks you for:<\/p>\n<ol>\n<li><strong>Assembly 1 folder.<\/strong> This folder should contain FASTA or multiFASTA files and must be inside assembly collection folder. Optionally, it could contain Bismark Bowtie2 indexes.<\/li>\n<li><strong>Adapter file.<\/strong> This file must be a multiFASTA file inside adapter collection.<\/li>\n<li><strong>Input data folder(s).<\/strong>\u00a0Each of these folders should contain all the input datasets of a sample in SRA, FASTQ, SAM or BAM format (all files must be in the same format) and must be inside the root input folder.<\/li>\n<\/ol>\n<p>If you want to use a second assembly, launch MethFlow as follow:<\/p>\n<pre style=\"text-align: center;\"><strong>MethFlow \u2010\u2010assembly2<\/strong><\/pre>\n<p>In this case, MethFlow asks you for:<\/p>\n<ol>\n<li><strong>Assembly 1 folder.<\/strong> This folder should contain FASTA or multiFASTA files and must be inside assembly collection folder. Optionally, it could contain Bismark Bowtie2 indexes.<\/li>\n<li><strong>Assembly 2 folder.<\/strong> This folder should contain FASTA or multiFASTA files and must be inside assembly collection folder. Optionally, it could contain Bismark Bowtie2 indexes.<\/li>\n<li><strong>What type of reads you want to use against the assembly 2:<\/strong> multiple-mapped reads, unmapped reads or both.<\/li>\n<li><strong>Adapter file.<\/strong> This file must be a multiFASTA file inside adapter collection.<\/li>\n<li><strong>Input data folder(s).<\/strong>\u00a0Each of these folders should contain all the input datasets of a sample in SRA, FASTQ, SAM or BAM format (all files must be in the same format) and must be inside the root input folder.<\/li>\n<\/ol>\n<p><span id=\"ouHighlight__0_13TO0_11\" class=\"\">Additionally<\/span><span id=\"noHighlight_0.8676234614908152\">,<\/span><span id=\"noHighlight_0.6976494981161827\"> <\/span><span id=\"ouHighlight__16_18TO14_16\" class=\"\">use<\/span><span id=\"noHighlight_0.0906570260582924\"> <\/span><span id=\"ouHighlight__20_22TO18_20\">the<\/span><span id=\"noHighlight_0.5782996075823763\"> <\/span><span id=\"ouHighlight__24_31TO22_28\">options<\/span><span id=\"noHighlight_0.14885440070328393\">\u00a0<strong>\u2010\u2010enable_api<\/strong><\/span><span id=\"noHighlight_0.11344022272910004\">\u00a0<\/span><span id=\"ouHighlight__36_36TO34_36\">and<\/span><span id=\"noHighlight_0.8178921835612691\"> <strong>\u2010\u2010enable_diffmeth<\/strong><\/span><span id=\"noHighlight_0.5483406972786837\">\u00a0<\/span><span id=\"ouHighlight__41_44TO42_43\">to<\/span><span id=\"noHighlight_0.7456307411930678\"> <\/span><span id=\"ouHighlight__46_52TO45_52\">activate<\/span><span id=\"noHighlight_0.6097980775447114\">\u00a0NGSmethDB API client<\/span><span id=\"noHighlight_0.19534596049887143\">\u00a0<\/span><span id=\"ouHighlight__56_56TO56_58\">and<\/span><span id=\"noHighlight_0.3839531351818044\">\u00a0differential methylation analysis functionalities<\/span><span id=\"ouHighlight__58_59TO60_61\">,<\/span><span id=\"noHighlight_0.714314756293355\"> <\/span><span id=\"ouHighlight__61_75TO63_74\">respectively<\/span><span id=\"noHighlight_0.6297298994401557\">.<\/span><span id=\"noHighlight_0.5849362428588822\"> <\/span><span id=\"ouHighlight__78_85TO77_84\">MethFlow asks\u00a0<\/span><span id=\"noHighlight_0.8807107168266013\"><\/span><span id=\"ouHighlight__87_88TO86_88\">you<\/span><span id=\"noHighlight_0.9520375869905775\"> <\/span><span id=\"ouHighlight__101_103TO97_100\">what<\/span><span id=\"noHighlight_0.4112756769424726\"> <\/span><span id=\"ouHighlight__105_112TO102_108\" class=\"\">samples<\/span><span id=\"noHighlight_0.9787954657415041\">\u00a0you want to\u00a0<\/span><span id=\"ouHighlight__114_122TO110_117\" class=\"\">download<\/span><span id=\"noHighlight_0.5888984200260194\">\u00a0from<\/span><span id=\"noHighlight_0.2157172174708979\">\u00a0<\/span><span id=\"ouHighlight__127_128TO122_124\">the<\/span><span id=\"noHighlight_0.33090849979118264\"> <\/span><span id=\"ouHighlight__130_138TO126_134\">NGSmethDB<\/span><span id=\"noHighlight_0.7411572490619205\"> <\/span><span id=\"ouHighlight__140_140TO136_138\">and<\/span><span id=\"noHighlight_0.14002499373866995\">\u00a0what<\/span><span id=\"noHighlight_0.5886416442989675\">\u00a0<\/span><span id=\"ouHighlight__146_153TO145_151\">samples<\/span><span id=\"noHighlight_0.9673121861636826\"> <\/span><span id=\"ouHighlight__155_162TO153_159\">compare\u00a0<\/span><span id=\"noHighlight_0.4510468058318138\"><\/span><span id=\"ouHighlight__164_165TO161_162\">in<\/span><span id=\"noHighlight_0.9755744369350245\"> <\/span><span id=\"ouHighlight__167_168TO164_166\" class=\"\">the\u00a0<\/span><span id=\"ouHighlight__193_203TO192_203\" class=\"\">differential<span id=\"noHighlight_0.2595808121142329\">\u00a0<\/span><span id=\"ouHighlight__182_191TO180_190\" class=\"\">methylation<\/span><span id=\"noHighlight_0.7804256309098638\"> <\/span>analysis<\/span><span id=\"noHighlight_0.9979496553751572\">.<\/span><\/p>\n<h3>A look at the output<\/h3>\n<ul>\n<li><strong>Intermediates folder:<\/strong> <span id=\"ouHighlight__14_18TO14_16\" class=\"\">all<\/span><span id=\"noHighlight_0.07545820414138715\"> <\/span><span id=\"ouHighlight__20_22TO18_20\">the<\/span><span id=\"noHighlight_0.9153987486654813\"> <\/span><span id=\"ouHighlight__33_43TO28_39\">intermediate files<\/span><span id=\"noHighlight_0.1286313150550824\"> <\/span><span id=\"ouHighlight__45_53TO41_49\" class=\"\">generated<\/span><span id=\"noHighlight_0.9301825002908815\"> <\/span><span id=\"ouHighlight__55_61TO51_56\" class=\"\">during<\/span><span id=\"noHighlight_0.08993821523127776\"> <\/span><span id=\"ouHighlight__63_64TO58_60\" class=\"\">the<\/span><span id=\"noHighlight_0.6872673620335985\"> <\/span><span id=\"ouHighlight__66_73TO62_69\" class=\"\">analysis<\/span><span id=\"noHighlight_0.22142384692696315\">.<\/span><strong>\u00a0<\/strong>It\u00a0<span id=\"ouHighlight__5_12TO5_12\" class=\"\">contains a\u00a0folder for each analyzed sample.<\/span><\/li>\n<li><strong>Plots folder:<\/strong><span id=\"noHighlight_0.7301467444298817\">\u00a0<\/span><span id=\"ouHighlight__18_22TO16_20\" class=\"\">plots<\/span><span id=\"noHighlight_0.5475378065103533\"> <\/span><span id=\"ouHighlight__24_32TO22_30\" class=\"\">generated<\/span><span id=\"noHighlight_0.02068600755891703\"> <\/span><span id=\"ouHighlight__34_36TO32_33\" class=\"\">by<\/span><span id=\"noHighlight_0.9507322942728866\"> <\/span><span id=\"ouHighlight__38_43TO35_40\" class=\"\">FastQC<\/span><span id=\"noHighlight_0.413405703844135\">,<\/span><span id=\"noHighlight_0.6919898626085998\"> <\/span><span id=\"ouHighlight__46_50TO43_47\" class=\"\">BSeQC<\/span><span id=\"noHighlight_0.06013854719396283\"> <\/span><span id=\"ouHighlight__52_52TO49_51\" class=\"\">and<\/span><span id=\"noHighlight_0.21523532471035267\"> <\/span><span id=\"ouHighlight__54_62TO53_61\" class=\"\">methylKit<\/span><span id=\"noHighlight_0.9295557173953668\">.<strong>\u00a0<\/strong>It\u00a0<span id=\"ouHighlight__5_12TO5_12\" class=\"\">contains a folder for each analyzed sample.<\/span><\/span><\/li>\n<li><strong>Meth folder:<\/strong><span id=\"noHighlight_0.5578221021297478\">\u00a0all\u00a0<\/span><span id=\"ouHighlight__27_36TO16_26\" class=\"\">methylation<\/span><span id=\"noHighlight_0.4938989405898653\"> <\/span><span id=\"ouHighlight__18_22TO28_31\" class=\"\">maps<\/span><span id=\"noHighlight_0.38932746822653286\"> <\/span><span id=\"ouHighlight__38_47TO33_42\" class=\"\">calculated<\/span><span id=\"noHighlight_0.3796660264738987\">\u00a0<\/span><span id=\"ouHighlight__49_51TO44_45\">by<\/span><span id=\"noHighlight_0.39273337788664775\"> <\/span><span id=\"ouHighlight__53_60TO47_54\" class=\"\">MethFlow<\/span><span id=\"noHighlight_0.8177048184320586\">.<\/span><span id=\"noHighlight_0.616119880618601\"> It\u00a0<span id=\"ouHighlight__5_12TO5_12\" class=\"\">contains a folder for each analyzed sample.<\/span><\/span><\/li>\n<\/ul>\n<p>If you use the NGSmethDB API client functionality, each of the downloaded methylation maps will be stored in a folder inside the Meth folder.<\/p>\n<p>If you use the differential methylation analysis functionality, you will have an additional output folder:<\/p>\n<ul>\n<li><strong>Diffmeth folder:<\/strong>\u00a0all<span id=\"noHighlight_0.8250479858252433\">\u00a0<\/span><span id=\"ouHighlight__38_48TO16_27\" class=\"\">differential<\/span><span id=\"noHighlight_0.45999008923868123\"> <\/span><span id=\"ouHighlight__27_36TO29_39\" class=\"\">methylation<\/span><span id=\"noHighlight_0.5553820625838168\"> <\/span><span id=\"ouHighlight__18_22TO41_44\">maps<\/span><span id=\"noHighlight_0.19512951934003575\"> <\/span><span id=\"ouHighlight__50_59TO46_55\">calculated<\/span><span id=\"noHighlight_0.28837372236284753\"> <\/span><span id=\"ouHighlight__61_63TO57_58\">by<\/span><span id=\"noHighlight_0.24902246054852317\"> <\/span><span id=\"ouHighlight__65_72TO60_67\">MethFlow<span id=\"noHighlight_0.8177048184320586\">.<\/span><span id=\"noHighlight_0.616119880618601\"> It\u00a0<span id=\"ouHighlight__5_12TO5_12\" class=\"\">contains a folder for each pair of samples used in the differential methylation analysis.<\/span><\/span><\/span><\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"<p>Quick start is thought to try MethFlow without going into details and using the MethFlow virtual machine. For further information, see\u00a0the\u00a0reference manual. Install MethFlowVM Install VirtualBox. Install VirtualBox Extension Pack. Download MethFlowVM\u00a0(mirror). Import MethFlowVM\u00a0to VirtualBox by double-clicking. Optional: add a<span class=\"ellipsis\">&hellip;<\/span><\/p>\n<div class=\"read-more\"><a href=\"https:\/\/bioinfo2.ugr.es\/MethFlow\/quick-start\/\">Read more &#8250;<\/a><\/div>\n<p><!-- end of .read-more --><\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/pages\/2192"}],"collection":[{"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/comments?post=2192"}],"version-history":[{"count":5,"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/pages\/2192\/revisions"}],"predecessor-version":[{"id":2315,"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/pages\/2192\/revisions\/2315"}],"wp:attachment":[{"href":"https:\/\/bioinfo2.ugr.es\/MethFlow\/wp-json\/wp\/v2\/media?parent=2192"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}