This is the new server for GMOD.org. Please let us know if you notice anything weird while it's getting broken in.

WebApollo Tutorial 2012

From GMOD
Jump to: navigation, search

This WebApollo tutorial was presented by Ed Lee at the 2012 GMOD Summer School.

To follow along with the tutorial, you will need to use AMI ID: ami-a1de69c8, name: GMOD 2012 start day 3, available in the US East (N. Virginia) region. See the GMOD Cloud Tutorial for information on how to get this AMI.

Please note that WebApollo was not officially released when this session was taught, and this tutorial was run using a beta version of the tool.

Introduction

WebApollo Presentation


Using WebApollo

WebApollo is a web-based application, so the only requirement to use it is a web browser. It has been tested with Chrome, Firefox, and Safari. It currently does not support Internet Explorer.

A WebApollo demo with the Pythium data has been set up on the virtual machine. We'll use the demo to view WebApollo's functionality.

Important: before you load the demo, you'll need to do a few things:

$ cd /usr/local/tomcat/tomcat7/webapps/WebApolloDemo/jbrowse
$ sudo wget http://icebox.lbl.gov/webapollo/data/genome.css
$ sudo mv genome.css.1 genome.css

Next we need to modify data/bam_trackList.json. You'll want to modify the data_url and index_url elements and add a sourceUrl element.

This is how it currently looks:

       "data_url" : "http://ec2-##-##-##-##.compute-1.amazonaws.com:8080/WebApolloDemo/jbrowse/data/bam/scf1117875582023.bam",
       "index_url" : "http://ec2-##-##-##-##.compute-1.amazonaws.com:8080/WebApolloDemo/jbrowse/data/bam/scf1117875582023.bam.bai",

We want to change it to:

       "data_url" : "bam/scf1117875582023.bam",
       "index_url" : "bam/scf1117875582023.bam.bai",
       "sourceUrl" : "data/",

Point your browser to http://ec2-##-##-##-##.compute-1.amazonaws.com:8080/WebApolloDemo.
(e.g., http://ec2-184-73-92-243.compute-1.amazonaws.com:8080/WebApolloDemo).

WebApollo login page

The user name and password are both demo.

WebApollo main options

Click on the Edit annotations button.

WebApollo reference sequence selection

We only have one contig to work with. Click on the Edit button.

  • Annotation track
  • Add evidence tracks (maker, snap_masked, blastn, blastx, est2genome, protein2genome, BAM alignments)
  • Moving around the contig
  • Go to region scf1117875582023:629476..633770
  • Selection
    • Edge matching
  • Create annotation
    • Drag-n-drop
  • Delete exon (notice the change in CDS)
  • Add exon
  • Non-canonical splice sites (GT / AG)
  • Zoom to base
  • DNA track
    • Highlighting
    • 6-frame translation
  • DNA sequence insertion, deletion, substitution
  • Zoom back out
  • Change exon boundaries
  • Merge (exons, transcripts)
  • Split (exons, transcripts)
  • Make intron
  • Duplicate
  • Set translation start
  • Realtime client updating
  • Undo / Redo
  • Comments
  • DBXrefs
  • Get sequence
  • Sequence searching

Installing WebApollo

Server operating system

Any Unix like system (e.g., Unix, Linux, Mac OS X)

Prerequisites

  • Servlet container (must support servlet spec 3.0+) [officially supported: Tomcat 7]
  • Relational Database Management System [officially supported: PostgreSQL]
  • User database helper scripts
    • Perl modules
      • DBI
      • DBD module for your specific RDBMS [officially supported: DBD::Pg for PostgresSQL]
  • Data generation pipeline (see JBrowse prerequisites for more information on its prerequisites)
  • Sequence search (optional)
    • Blat (along with a configured search database)

Note: All prerequisites have already been installed on the course's machine

Installation

The installation steps will be done in the command line. SSH into your AWS machine.

Uncompress the WebApollo.tgz tarball.

$ cd
$ tar -xvzf WebApollo.tgz

User database

First we’ll need to create a database that will contain user permission information. WebApollo uses this database to determine who can access and edit annotations for a given sequence. We'll create the user web_apollo_users_admin to be the owner of the database with the password web_apollo_users_admin (enter it when prompted). The user will not be a superuser nor will it be able to create new roles. But it will be able to create databases.

$ createuser -P web_apollo_users_admin
Enter password for new role:
Enter it again:
Shall the new role be a superuser? (y/n) n
Shall the new role be allowed to create databases? (y/n) y
Shall the new role be allowed to create more new roles? (y/n) n

Let's now create the database. We can call it whatever we want, so let's call it web_apollo_users.

$ createdb -U web_apollo_users_admin web_apollo_users

Now that the database is created, we need to load the schema to it.

$ cd ~/WebApollo/tools/user
$ psql -U web_apollo_users_admin web_apollo_users < user_database_postgresql.sql

Now the user database has been setup.

Let's populate the database.

First we’ll create an user with access to WebApollo. We’ll use the add_user.pl script in ~/WebApollo/tools/user. Let’s create an user named web_apollo_admin with the password web_apollo_admin.

$ ./add_user.pl -D web_apollo_users -U web_apollo_users_admin -P web_apollo_users_admin \
   -u web_apollo_admin -p web_apollo_admin

Next we’ll add the genomic sequence ids for our organism in the database. We’ll use the add_tracks.pl script in the same directory. We need to generate a file of genomic sequence ids for the script. For convenience, there’s a script called extract_seqids_from_fasta.pl in the same directory which will go through a FASTA file and extract all the ids from the deflines. We’ll use the data from MAKER. Let’s first create the list of genomic sequence ids. We'll store it in ~/scratch/seqids.txt. We’ll want to add the prefix “Annotations-” to each identifier.

$ mkdir ~/scratch
$ ./extract_seqids_from_fasta.pl -p Annotations- -i ~/maker_output/scf1117875582023.fa \
   -o ~/scratch/seqids.txt

Now we’ll add those ids to the user database.

$ ./add_tracks.pl -D web_apollo_users -U web_apollo_users_admin -P web_apollo_users_admin \
   -t ~/scratch/seqids.txt

Now that we have an user created and the genomic sequence ids loaded, we’ll need to give the user permissions to access the sequence. We’ll have the user have read, write, and user manager permissions. We’ll use the set_track_permissions.pl script in the same directory. We’ll need to provide the script a list of genomic sequence ids, like in the previous step.

$ ./set_track_permissions.pl -D web_apollo_users -U web_apollo_users_admin \
   -P web_apollo_users_admin -u web_apollo_admin -t ~/scratch/seqids.txt -r -w -m

We’re all done setting up the user database.

Note that we’re only using a subset of the options for all the scripts mentioned above. You can get more detailed information on any given script (and other available options) using the “-h” or “--help” flag when running the script.

Deploy servlet

We need to deploy the WAR file in the war directory from the unpacked tarball. We need to go into the webapps directory in our Tomcat installation. Tomcat's installed in /usr/local/tomcat/tomcat7/.

$ cd /usr/local/tomcat/tomcat7/webapps 

Next we need to create the directory that will contain the application. Note that you'll need to use sudo for this, since only root has access to the directory.

$ sudo mkdir WebApollo

Now we'll go into the newly created directory and unpack the WAR file into it.

$ cd WebApollo
$ sudo jar -xvf ~/WebApollo/war/WebApollo.war


That’s it! We’re done installing WebApollo. Now we need to move on to configuring the application.

Configuration

Most configuration files will reside in /usr/local/tomcat/tomcat7/webapps/WebApollo/config. We’ll need to configure a number of things before we can get WebApollo up and running.

Main configuration

The main configuration is stored in /usr/local/tomcat/tomcat7/webapps/WebApollo/config/config.xml. Let’s take a look at the file.

<?xml version="1.0" encoding="UTF-8"?>
<server_configuration>
 
	<!-- mapping configuration for GBOL data structures -->
	<gbol_mapping>/config/mapping.xml</gbol_mapping>
 
	<!-- directory where JE database will be created -->
	<datastore_directory>ENTER_DATASTORE_DIRECTORY_HERE</datastore_directory>
 
	<!-- minimum size for introns created -->
	<default_minimum_intron_size>1</default_minimum_intron_size>
 
	<!-- size of history for each feature - setting to 0 means unlimited history -->
	<history_size>0</history_size>
 
	<!-- overlapping strategy for adding transcripts to genes -->
	<overlapper_class>org.bbop.apollo.web.overlap.OrfOverlapper</overlapper_class>
 
	<!-- class for comparing track names (used for sorting in lists) -->
	<track_name_comparator_class>org.bbop.apollo.web.track.DefaultTrackNameComparator</track_name_comparator_class>
 
	<!-- user authentication/permission configuration -->
	<user>
 
		<!-- database configuration -->
		<database>
 
			<!-- driver for user database -->
			<driver>org.postgresql.Driver</driver>
 
			<!-- JDBC URL for user database -->
			<url>ENTER_USER_DATABASE_JDBC_URL</url>
 
			<!-- username for user database -->
			<username>ENTER_USER_DATABASE_USERNAME</username>
 
			<!-- password for user database -->
			<password>ENTER_USER_DATABASE_PASSWORD</password>
 
		</database>
 
		<!-- class for generating user authentication page (login page) -->
		<authentication_class>org.bbop.apollo.web.user.localdb.LocalDbUserAuthentication</authentication_class>
 
	</user>
 
	<tracks>
 
		<!-- path to JBrowse refSeqs.json file -->
		<refseqs>ENTER_PATH_TO_REFSEQS_JSON_FILE</refseqs>
 
		<!-- annotation track name the current convention is to append
		the genomic region id to the the name of the annotation track
		e.g., if the annotation track is called "Annotations" and the
		genomic region is chr2L, the track name will be
		"Annotations-chr2L".-->
		<annotation_track_name>Annotations</annotation_track_name>
 
	 	<!-- organism being annotated -->
		<organism>ENTER_ORGANISM</organism>
 
		<!-- CV term for the genomic sequences - should be in the form
		of "CV:TERM".  This applies to all sequences -->
		<sequence_type>ENTER_CVTERM_FOR_SEQUENCE</sequence_type>
 
	</tracks>
 
	<!-- path to file containing canned comments XML -->
	<canned_comments>/config/canned_comments.xml</canned_comments>
 
	<!-- tool to be used for sequence searching.  This is optional.
	If this is not setup, WebApollo will not have sequence search support -->
	<sequence_search_tool>
 
		<!-- class for handling search -->
		<class>org.bbop.apollo.tools.seq.search.blat.BlatCommandLine</class>
 
		<!-- configuration for search tool -->
		<config>/config/blat_config.xml</config>
 
	</sequence_search_tool>
 
</server_configuration>

Let’s look through each element in more detail with values filled in.

	<!-- mapping configuration for GBOL data structures -->
	<gbol_mapping>/config/mapping.xml</gbol_mapping>

File that contains type mappings used by the underlying data model. It’s best not to change the default option.

	<!-- directory where JE database will be created -->
	<datastore_directory>/data/dataHome/web_apollo/annotations</datastore_directory>

Directory where user generated annotations will be stored. The data is stored using Berkeley DB. We’ll use /data/dataHome/web_apollo/annotations.

	<!-- minimum size for introns created -->
	<default_minimum_intron_size>1</default_minimum_intron_size>

Minimum length of intron to be created when using the “Make intron” operation. The operation will try to make the shortest intron that’s at least as long as this parameter. So if you set it to a value of “40”, then all calculated introns will be at least of length 40.

	<!-- size of history for each feature - setting to 0 means unlimited history -->
	<history_size>0</history_size>

The size of your history stack, meaning how many “Undo/Redo” steps you can do. The larger the number, the larger the storage space needed. Setting it to “0” makes it to that there’s no limit.

	<!-- overlapping strategy for adding transcripts to genes -->
	<overlapper_class>org.bbop.apollo.web.overlap.OrfOverlapper</overlapper_class>

Defines the strategy to be used for deciding whether overlapping transcripts should be considered splice variants to the same gene. This points to a Java class implementing the org.bbop.apollo.overlap.Overlapper interface. This allows you to create your own custom overlapping strategy should the need arise. Currently available options are:

  • org.bbop.apollo.web.overlap.NoOverlapper
    • No transcripts should be considered splice variants, regardless of overlap.
  • org.bbop.apollo.web.overlap.SimpleOverlapper
    • Any overlapping of transcripts will cause them to be part of the same gene
  • org.bbop.apollo.web.overlap.OrfOverlapper
    • Only transcripts that overlap within the coding region and within frame are considered part of the same gene
	<!-- class for comparing track names (used for sorting in lists) -->
	<track_name_comparator_class>org.bbop.apollo.web.track.DefaultTrackNameComparator</track_name_comparator_class>

Defines how to compare genomic sequence names for sorting purposes in the genomic region selection list. Points to a class implementing the org.bbop.apollo.web.track.TrackNameComparator interface. You can implement your own class to allow whatever sorting you’d like for your own organism. This doesn't make much of a difference in our case since we're only dealing with one genomic region. The only available implementation is:

  • org.bbop.apollo.web.track.DefaultTrackNameComparator
    • Sorts genomic sequence names lexicographically

Let’s take look at the “user” element, which handles configuration for user authentication and permission handling.

	<!-- user authentication/permission configuration -->
	<user>
 
		<!-- database configuration -->
		<database>
 
			<!-- driver for user database -->
			<driver>org.postgresql.Driver</driver>
 
			<!-- JDBC URL for user database -->
			<url>ENTER_USER_DATABASE_JDBC_URL</url>
 
			<!-- username for user database -->
			<username>ENTER_USER_DATABASE_USERNAME</username>
 
			<!-- password for user database -->
			<password>ENTER_USER_DATABASE_PASSWORD</password>
 
		</database>
 
		<!-- class for generating user authentication page (login page) -->
		<authentication_class>org.bbop.apollo.web.user.localdb.LocalDbUserAuthentication</authentication_class>
 
	</user>

Let’s first look at the database element that defines the database that will handle user permissions (which we created previously).

			<!-- driver for user database -->
			<driver>org.postgresql.Driver</driver>

This should point the JDBC driver for communicating with the database. We’re using a PostgreSQL driver since that’s the database we’re using for user permission management.

			<!-- JDBC URL for user database -->
			<url>jdbc:postgresql://localhost/web_apollo_users</url>

JDBC URL to the user permission database. We'll use jdbc:postgresql://localhost/web_apollo_users since the database is running in the same server as the annotation editing engine and we named the database web_apollo_users.

			<!-- username for user database -->
			<username>web_apollo_users_admin</username>

User name that has read/write access to the user database. The user with access to the user database has the user name web_apollo_users_admin.

			<!-- password for user database -->
			<password>web_apollo_users_admin</password>

Password to access user database. The user with access to the user database has the password </tt>web_apollo_users_admin</tt>.

Now let’s look at the other elements in the user element.

			<!-- class for generating user authentication page (login page) -->
			<authentication_class>org.bbop.apollo.web.user.localdb.LocalDbUserAuthentication</authentication_class>

Defines how user authentication is handled. This points to a class implementing the org.bbop.apollo.web.user.UserAuthentication interface. This allows you to implement any type of authentication you’d like (e.g., LDAP). Currently available options are:

  • org.bbop.apollo.web.user.localdb.LocalDbUserAuthentication
    • Uses the user permission database to also store authentication information, meaning it stores user passwords in the database
  • org.bbop.apollo.web.user.browserid.BrowserIdUserAuthentication
    • Uses Mozilla’s BrowserID service for authentication. This has the benefits of offloading all authentication security to Mozilla and allows one account to have access to multiple resources (as long as they have BrowserID support). Being that the service is provided through Mozilla, it will require users to create a BrowserID account

Now let’s look at the configuration for accessing the annotation tracks for the genomic sequences.

	<tracks>
 
		<!-- path to JBrowse refSeqs.json file -->
		<refseqs>ENTER_PATH_TO_REFSEQS_JSON_FILE</refseqs>
 
		<!-- annotation track name the current convention is to append
		the genomic region id to the the name of the annotation track
		e.g., if the annotation track is called "Annotations" and the
		genomic region is chr2L, the track name will be
		"Annotations-chr2L".-->
		<annotation_track_name>Annotations</annotation_track_name>
 
	 	<!-- organism being annotated -->
		<organism>ENTER_ORGANISM</organism>
 
		<!-- CV term for the genomic sequences - should be in the form
		of "CV:TERM".  This applies to all sequences -->
		<sequence_type>ENTER_CVTERM_FOR_SEQUENCE</sequence_type>
 
	</tracks>

Let’s look at each element individually.

		<!-- path to JBrowse refSeqs.json file -->
		<refseqs>/usr/local/tomcat/tomcat7/webapps/WebApollo/jbrowse/data/refSeqs.json</refseqs>

Location where the refSeqs.json file resides, which is created from the data generation pipeline (see the data generation section). By default, the JBrowse data needs to reside in /usr/local/tomcat/tomcat7/webapps/WebApollo/jbrowse/data. If you want the data to reside elsewhere, you’ll need to do configure your servlet container to handle the appropriate alias to jbrowse/data.

		<annotation_track_name>Annotations</annotation_track_name>

Name of the annotation track. Leave it as the default value of Annotations.

	 	<!-- organism being annotated -->
		<organism>Pythium ultimum</organism>

Scientific name of the organism being annotated (genus and species). We're annotating Pythium ultimum.

		<!-- CV term for the genomic sequences - should be in the form
		of "CV:TERM".  This applies to all sequences -->
		<sequence_type>sequence:contig</sequence_type>

The type for the genomic sequences. Should be in the form of CV:term. Our genomic sequences are of the type sequence:contig.

	<!-- path to file containing canned comments XML -->
	<canned_comments>/config/canned_comments.xml</canned_comments>

File that contains canned comments (predefined comments that will be available from a pull-down menu when creating comments). It’s best not to change the default option. See the canned comments section for details on configuring canned comments.

	<!-- tool to be used for sequence searching.  This is optional.
	If this is not setup, WebApollo will not have sequence search support -->
	<sequence_search_tool>
 
		<!-- class for handling search -->
		<class>org.bbop.apollo.tools.seq.search.blat.BlatCommandLine</class>
 
		<!-- configuration for search tool -->
		<config>/config/blat_config.xml</config>
 
	</sequence_search_tool>

Here’s the configuration for sequence search tools (allows searching your genomic sequences). WebApollo does not implement any search algorithms, but instead relies on different tools and resources to handle searching (this provides much more flexible search options). This is optional. If it’s not configured, WebApollo will not have sequence search support. There are two elements that need to be configured.

		<!-- class for handling search -->
		<class>org.bbop.apollo.tools.seq.search.blat.BlatCommandLine</class>

Should point to the class that will handle the search request. Searching is handled by classes that implement the org.bbop.apollo.tools.seq.search.SequenceSearchTool interface. This allows you to add support for your own favorite search tools (or resources). We currently only have support for command line Blat.

		<!-- configuration for search tool -->
		<config>/config/blat_config.xml</config>

File that contains the configuration for the searching plugin chosen. If you’re using Blat, you should not change this. If you’re using your own plugin, you’ll want to point this to the right configuration file (which will be dependent on your plugin). See the Blat section for details on configuring WebApollo to use Blat.

Canned comments

You can configure a set of predefined comments that will be available for users when adding comments through a dropdown menu. The configuration is stored in /usr/local/tomcat/tomcat7/webapps/WebApollo/config/canned_comments.xml. Let’s take a look at the configuration file.

<?xml version="1.0" encoding="UTF-8"?>
 
<canned_comments>
	<!-- one <comment> element per comment.
	it must contain the attribute "feature_type" that defines
	the type of feature this comment will apply to.
	must be be in the form of "cv:cvterm" (e.g., "sequence:gene")
 
	<comment feature_type="sequence:gene">This is a comment for sequence:gene</comment>
	-->
</canned_comments>

You’ll need one <comment> element for each predefined comment. The element needs to have a feature_type attribute in the form of cv:cvterm that this comment applies to. Let’s make a few comments for feature of type sequence:gene and sequence:transcript:

	<comment feature_type="sequence:gene">This is a comment for a gene</comment>
	<comment feature_type="sequence:gene">This is another comment for a gene</comment>
	<comment feature_type="sequence:transcript">This is a comment for a transcript</comment>

Search tools

As mentioned previously, WebApollo makes use of tools for sequence searching rather than employing its own search algorithm. The only currently supported tool is command line Blat.

Blat

You’ll need to have Blat installed and a search database with your genomic sequences available to make use of this feature. The configuration is stored in /usr/local/tomcat/tomcat7/webapps/WebApollo/config/blat_config.xml. Let’s take a look at the configuration file:

<?xml version="1.0" encoding="UTF-8"?>
 
<!-- configuration file for setting up command line Blat support -->
 
<blat_config>
 
	<!-- path to Blat binary →
	<blat_bin>ENTER_PATH_TO_BLAT_BINARY</blat_bin>
 
	<!-- path to where to put temporary data -->
	<tmp_dir>ENTER_PATH_FOR_TEMPORARY_DATA</tmp_dir>
 
	<!-- path to Blat database -->
 	<database>ENTER_PATH_TO_BLAT_DATABASE</database>
 
	<!-- any Blat options (directly passed to Blat) e.g., -minMatch -->
	<blat_options>ENTER_ANY_BLAT_OPTIONS</blat_options>
 
</blat_config>

Let’s look at each element with values filled in.

	<!-- path to Blat binary -->
	<blat_bin>/home/ubuntu/Software/blat/blat</blat_bin>

We need to point to the location where the Blat binary resides. We it installed in /usr/local/bin/blat.

	<!-- path to where to put temporary data -->
	<tmp_dir>/data/dataHome/web_apollo/blat/tmp</tmp_dir>

We need to point to the location where to store temporary files to be used in the Blat search. It can be set to whatever location you’d like. We’ll store the temporary data to /data/dataHome/web_apollo/tmp.

	<!-- path to Blat database -->
	<database>/data/dataHome/web_apollo/blat/db/pyu.2bit</database>

We need to point to the location of the search database to be used by Blat. We need point it to /data/dataHome/web_apollo/blat/db/pyu.2bit. We're using a 2bit database in this case, but you have options on how to generate your Blat database. See the Blat documentation for more information.

	<!-- any Blat options (directly passed to Blat) e.g., -minMatch -->
	<blat_options>-minScore=100 -minIdentity=60</blat_options>

Here we can configure any extra options to used by Blat. These options are passed verbatim to the program. In this example, we’re passing the -minScore parameter with a minimum score of 100 and the -minIdentity parameter with a value of 60 (60% identity). See the Blat documentation for information of all available options.

Data generation

The steps for generating data (in particular static data) are mostly similar to JBrowse data generation steps, with some extra steps required. Most of the scripts for data generation reside in /usr/local/tomcat/tomcat7/webapps/WebApollo/jbrowse/bin. Let's go into WebApollo's JBrowse directory.

$ cd /usr/local/tomcat/tomcat7/webapps/WebApollo/jbrowse

It will make things easier if we make sure that the scripts in the bin directory are executable. Note that you'll need to have root privilege to do so.

$ sudo chmod 755 bin/*

As mentioned previously, the data resides in the data directory by default. Let's create a symlink to /data/dataHome/web_apollo/jbrowse/data to be used as the data directory. This gives a lot of flexibility in allowing your WebApollo instance to easily point to a new data directory. Note that you'll need to have root privilege to do so.

$ sudo ln -sf /data/dataHome/web_apollo/jbrowse/data

Now that we have our data directory in JBrowse, we need to copy some files into it that are specific to WebApollo's JBrowse. We need to copy all of the contents from ~/WebApollo/json in our data directory.

$ cp ~/WebApollo/json/* data

The first thing we need to do before processing our evidence is to generate the reference sequence data to be used by JBrowse. We'll use the prepare-refseqs.pl script.

$ bin/prepare-refseqs.pl --fasta ~/maker_output/scf1117875582023.fa

We now have the DNA track setup. WebApollo's DNA track is modified from JBrowse's stock DNA track, so we need to modify the newly generated data/trackList.json file. This step IS VERY IMPORTANT, otherwise WebApollo will not work.

First, let's look at the stock generated data/trackList.json file.

{
  "tracks" : [
     {
        "chunkSize" : 20000,
        "urlTemplate" : "seq/{refseq}/",
        "type" : "SequenceTrack",
        "label" : "DNA",
        "key" : "DNA"
     }
  ],
  "formatVersion" : 1
}

We need to modify the track object. We need to modify the type from SequenceTrack to AnnotSequenceTrack, remove the urlTemplate , move the chunkSize element into the config member we'll be adding:

        "config" : {
           "chunkSize" : 20000,
           "residuesUrlTemplate" : "seq/{refseq}/",
           "urlTemplate" : "sequence_alterations_trackData.json",
           "style" : {
              "uniqueIdField" : "id",
              "className" : "{type}"
           },
           "compress" : 0,
           "type" : 1,
           "subfeatures" : 1
        }

After the modifications, the file should look as follow (you can just copy and paste it):

{
  "tracks" : [
     {
        "type" : "AnnotSequenceTrack",
        "label" : "DNA",
        "key" : "DNA",
        "config" : {
           "chunkSize" : 20000,
           "residuesUrlTemplate" : "seq/{refseq}/",
           "urlTemplate" : "sequence_alterations_trackData.json",
           "style" : {
              "uniqueIdField" : "id",
              "className" : "{type}"
           },
           "compress" : 0,
           "type" : 1,
           "subfeatures" : 1
        }
     }
  ],
  "formatVersion" : 1
}

We need to symlink the data/seq/refSeqs.json file in data/seq.

$ cd data
$ ln -sf seq/refSeqs.json
$ cd ..


Static data generation

Generating data from GFF3 works best by having a separate GFF3 per source type. The output from Maker puts all sources in the same GFF3. We need to split up the GFF3. We can use the split_gff_by_source.pl script in ~/WebApollo/tools/data to do so. We'll output the split GFF3 to some temporary directory (we'll use ~/scratch/split_gff).

$ mkdir -p ~/scratch/split_gff
$ ~/WebApollo/tools/data/split_gff_by_source.pl -i ~/maker_output/scf1117875582023.gff -d ~/scratch/split_gff

If we look at the contents of ~/scratch/split_gff, we can see we have the following files:

$ ls ~/scratch/split_gff
blastn.gff  est2genome.gff  protein2genome.gff  repeatrunner.gff
blastx.gff  maker.gff       repeatmasker.gff    snap_masked.gff

We need to process each file and create the appropriate tracks.

We'll start off with maker.gff. We need to handle that file a bit differently than the rest of the files since the GFF represents the features as gene, transcript, exons, and CDSs.

$ bin/flatfile-to-json.pl --gff ~/scratch/split_gff/maker.gff \
--arrowheadClass trellis-arrowhead --getSubfeatures \
--subfeatureClasses '{"wholeCDS": "null", "CDS":"maker-CDS", "UTR": "maker-UTR", "exon":"maker-exon"}' \
--cssClass maker-transcript --type mRNA --trackLabel maker

Now we need to convert the Maker track into WebApollo compatible JSON. We can use json_converter.sh in ~/WebApollo/tools/data to do so.

$ ~/WebApollo/tools/data/json_converter.sh -d data/tracks/maker
root track dir: data/tracks/maker
editing files in place
seq dir count: 1
done processing features, total scanned: 353

Lastly we need to modify the maker entry in data/trackList.json. Let's look at the stock entry.

     {
        "autocomplete" : "none",
        "style" : {
           "className" : "maker-transcript",
           "subfeatureClasses" : {
              "wholeCDS" : "null",
              "exon" : "maker-exon",
              "CDS" : "maker-CDS",
              "UTR" : "maker-UTR"
           },
           "arrowheadClass" : "trellis-arrowhead"
        },
        "key" : "maker",
        "phase" : null,
        "urlTemplate" : "tracks/maker/{refseq}/trackData.json",
        "compress" : 0,
        "label" : "maker",
        "type" : "FeatureTrack",
        "subfeatures" : 1
     }

We need to add "renderClassName" : "maker-transcript-render" to the style member. We also need to change type from FeatureTrack to DraggableFeatureTrack. This is how the updated entry should look like:

    {
        "autocomplete" : "none",
        "style" : {
           "renderClassName" : "maker-transcript-render",
           "className" : "maker-transcript",
           "subfeatureClasses" : {
              "wholeCDS" : "null",
              "exon" : "maker-exon",
              "CDS" : "maker-CDS",
              "UTR" : "maker-UTR"
           },
           "arrowheadClass" : "trellis-arrowhead"
        },
        "key" : "maker",
        "phase" : null,
        "urlTemplate" : "tracks/maker/{refseq}/trackData.json",
        "compress" : 0,
        "label" : "maker",
        "type" : "DraggableFeatureTrack",
        "subfeatures" : 1
     }

Now we need to process the other remaining GFF3 files. The entries in those are stored as "match/match_part", so they can all be handled in a similar fashion.

We'll start off with blastn as an example.

$ bin/flatfile-to-json.pl --gff ~/scratch/split_gff/blastn.gff \
--arrowheadClass trellis-arrowhead --getSubfeatures \
--subfeatureClasses '{"match_part": "blastn-alignment-part"}' \
--cssClass "blastn-alignment-match" --trackLabel blastn

Now we need to modify the blastn entry in data/trackList.json. Let's look at the stock entry.

     {
        "autocomplete" : "none",
        "style" : {
           "className" : "blastn-match",
           "subfeatureClasses" : {
              "match_part" : "blastn-part"
           },
           "arrowheadClass" : "trellis-arrowhead"
        },
        "key" : "blastn",
        "phase" : null,
        "urlTemplate" : "tracks/blastn/{refseq}/trackData.json",
        "compress" : 0,
        "label" : "blastn",
        "type" : "FeatureTrack",
        "subfeatures" : 1
     }

We need to add "renderClassName" : "blastn-alignment-render" to the style member. We also need to change type from FeatureTrack to DraggableFeatureTrack. This is how the updated entry should look like:

     {
        "autocomplete" : "none",
        "style" : {
           "renderClassName" : "blastn-alignment-render",
           "className" : "blastn-match",
           "subfeatureClasses" : {
              "match_part" : "blastn-part"
           },
           "arrowheadClass" : "trellis-arrowhead"
        },
        "key" : "blastn",
        "phase" : null,
        "urlTemplate" : "tracks/blastn/{refseq}/trackData.json",
        "compress" : 0,
        "label" : "blastn",
        "type" : "DraggableFeatureTrack",
        "subfeatures" : 1
     }

We need to follow the same steps for the remaining GFF3 files. It can be a bit tedious to do this for the remaining six files, so we can use a simple inline shell script to help us out. However, we'll still have to manually modify data/trackList.json for each new track. Don't worry if the script doesn't make sense, you can still process each file by hand. For now you can just copy and paste the script into your terminal.

$ for i in $(ls ~/scratch/split_gff/*.gff | grep -v maker);
do j=$(basename $i); j=${j/.gff/};
echo "Processing $j" && bin/flatfile-to-json.pl --gff $i --arrowheadClass trellis-arrowhead \
--getSubfeatures --subfeatureClasses "{\"match_part\": \"$j-alignment-part\"}" \
--cssClass "$j-alignment-match" --trackLabel $j; done

IMPORTANT!

Remember that we still need to add "renderClassName" : "$type-alignment-render" to the style member, where $type is the track name. We also need to change type from FeatureTrack to DraggableFeatureTrack.

Now let's look how to configure BAM support. WebApollo has native support for BAM, so extra processing of the data is required.

First we'll copy the BAM data into the WebApollo data directory. We'll put it in the data/bam directory. Keep in mind that this BAM data was randomly generated, so there's really no biological meaning to it. We only created it to show BAM support.

$ mkdir data/bam
$ cp ~/maker_output/*.bam* data/bam

Now we need to edit the data/bam_trackList.json file. Let's look at the file.

{
  "tracks" : [
     {
       "type" : "BamFeatureTrack",
       "label" : "ENTER_LABEL",
       "data_url" : "ENTER_URL_TO_BAM_FILE",
       "index_url" : "ENTER_URL_TO_BAI_FILE",
        "config" : {
           "autocomplete" : "none",
           "style" : {
              "className" : "bam",
              "subfeatureClasses" : {
                  "M": "cigarM",
                  "D": "cigarD",
                  "N": "cigarN",
                  "=": "cigarEQ",
                  "E": "cigarEQ",
                  "X": "cigarX",
                  "I": "cigarI"
              },
              "arrowheadClass" : null
           },
           "key" : "ENTER_KEY",
           "compress" : 0,
           "type" : 1,
           "subfeatures" : 1
        }
     }
   ],
  "formatVersion" : 1
}

We need to modify the following elements: label, data_url, index_url, and key.

Let's look at each value with their values filled in.

       "label" : "simulated_bam",

The internal name for this BAM track.

       "data_url" : "bam/scf1117875582023.bam",

URL to BAM file.

       "index_url" : "bam/scf1117875582023.bam.bai",

URL to BAM index file.

       "sourceUrl" : "data/",

Add sourceUrl under index_url.

You can point to the BAM/BAI file in any server by providing an absolute URL. This is a nice feature since BAM files can get rather large, so you don't need to create a local copy on your own server.

           "key" : "simulated BAM",

The external, human-readable label seen on the BAM track.

Here's how the updated JSON should look like.

{
  "tracks" : [
     {
       "type" : "BamFeatureTrack",
       "label" : "simulated_bam", 
       "data_url" : "bam/scf1117875582023.bam", 
       "index_url" : "bam/scf1117875582023.bam.bai",
       "sourceUrl" : "data/",
        "config" : {
           "autocomplete" : "none",
           "style" : {
              "className" : "bam",
              "subfeatureClasses" : {
                  "M": "cigarM",
                  "D": "cigarD",
                  "N": "cigarN",
                  "=": "cigarEQ",
                  "E": "cigarEQ",
                  "X": "cigarX",
                  "I": "cigarI"
              },
              "arrowheadClass" : null
           },
           "key" : "simulated BAM",
           "compress" : 0,
           "type" : 1,
           "subfeatures" : 1
        }
     }
  ],
  "formatVersion" : 1
}

You should now have a simulated BAM track now available.

You can download the updated trackList.json file here (you'll need to put the file in your virtual machine and unzip it in /usr/local/tomcat/tomcat7/webapps/WebApollo/jbrowse/data):
web_apollo_track_list_json.zip

Alternatively, you can just copy the WebApolloDemo trackList.json into WebApollo (since they're both using the same dataset).

$ cd /usr/local/tomcat/tomcat7/webapps/WebApollo/jbrowse/data
$ cp /usr/local/tomcat/tomcat7/webapps/WebApolloDemo/jbrowse/data/trackList.json .

Congratulations, you're done configuring WebApollo.

Let's test out our installation. Point your browser to http://ec2-##-##-##-##.compute-1.amazonaws.com:8080/WebApollo
(e.g., http://ec2-184-73-92-243.compute-1.amazonaws.com:8080/WebApollo).

WebApollo login page

The user name and password are both web_apollo_admin as we configured earlier.

WebApollo main options

Click on the Edit annotations button.

WebApollo reference sequence selection

We only see one reference sequence to annotate since we're only working with one contig. Click on the Edit button.

Now have fun annotating!!!