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SeqShop: Variant Calling and Filtering for SNPs Practical, December 2014 (view source)

Revision as of 19:12, 30 March 2015

214 bytes added ,  19:12, 30 March 2015
→‎Sequnce Alignment Files: BAM Files
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==Introduction==
 
==Introduction==
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See the [[Media:SeqShop - GotCloud snpcall.pdf|introductory slides]] for an intro to this tutorial.
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Main Workshop wiki page: [[SeqShop: December 2014]]
    +
See the [[Media:Dec2014 SeqShop - GotCloud snpcall.pdf|introductory slides]] for an intro to this tutorial.
    
== Goals of This Session ==
 
== Goals of This Session ==
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== Setup in person at the SeqShop Workshop ==
 
== Setup in person at the SeqShop Workshop ==
 
''This section is specifically for the SeqShop Workshop computers.''
 
''This section is specifically for the SeqShop Workshop computers.''
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<div class="mw-collapsible" style="width:600px">
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<div class="mw-collapsible mw-collapsed" style="width:600px">
 
''If you are not running during the SeqShop Workshop, please skip this section.''
 
''If you are not running during the SeqShop Workshop, please skip this section.''
 
<div class="mw-collapsible-content">
 
<div class="mw-collapsible-content">
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* Setup an output directory
 
* Setup an output directory
 
** It will leave your output directory from the previous tutorial in tact.
 
** It will leave your output directory from the previous tutorial in tact.
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  source /home/mktrost/seqshop/setup.txt
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  source /net/seqshop-server/home/mktrost/seqshop/setup.txt
 
* You won't see any output after running <code>source</code>
 
* You won't see any output after running <code>source</code>
 
** It silently sets up your environment
 
** It silently sets up your environment
 
** If you want to view the detail of the setup, type
 
** If you want to view the detail of the setup, type
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  less /home/mktrost/seqshop/setup.txt
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  less /net/seqshop-server/home/mktrost/seqshop/setup.txt
 
and press 'q' to finish.
 
and press 'q' to finish.
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== Setup when running on your own outside of the SeqShop Workshop ==
 
== Setup when running on your own outside of the SeqShop Workshop ==
 
''This section is specifically for running on your own outside of the SeqShop Workshop.''
 
''This section is specifically for running on your own outside of the SeqShop Workshop.''
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<div class="mw-collapsible mw-collapsed" style="width:600px">
+
<div class="mw-collapsible" style="width:600px">
 
''If you are running during the SeqShop Workshop, please skip this section.''
 
''If you are running during the SeqShop Workshop, please skip this section.''
 
<div class="mw-collapsible-content">
 
<div class="mw-collapsible-content">
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{{SeqShopRemoteEnv}}
 
{{SeqShopRemoteEnv}}
 +
</div>
 +
</div>
 +
    
== Examining GotCloud SnpCall Input files ==
 
== Examining GotCloud SnpCall Input files ==
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=== Sequnce Alignment Files: BAM Files ===
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=== Sequence Alignment Files: BAM Files ===
 
Per sample BAM files contain sequence reads that are mapped to positions in the genome.
 
Per sample BAM files contain sequence reads that are mapped to positions in the genome.
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<div class="mw-collapsible-content">
 
<div class="mw-collapsible-content">
 
<ul>
 
<ul>
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<li>/home/YourUserName/out/bams/HG00640.recal.bam</li>
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<li>/net/seqshop-server/home/YourUserName/out/bams/HG00640.recal.bam</li>
 
[[File:BamindexNew.png|500px]]
 
[[File:BamindexNew.png|500px]]
 
</div>
 
</div>
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<li>That's ok, we will use the <code>--base_prefix ${SS}</code> command-line option to prefix the BAM paths</li>
 
<li>That's ok, we will use the <code>--base_prefix ${SS}</code> command-line option to prefix the BAM paths</li>
 
<li>Alternatively, we could have set BAM_PREFIX in <code>gotcloud.conf</code> to the path to the BAMs
 
<li>Alternatively, we could have set BAM_PREFIX in <code>gotcloud.conf</code> to the path to the BAMs
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<pre>BAM_PREFIX = /home/username/seqshop/example</pre> </li>
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<pre>BAM_PREFIX = /net/seqshop-server/home/mktrost/seqshop/example</pre> </li>
 
<ul>
 
<ul>
 
<li>NOTE: the conf file can't interpret ${SS} environment variables or '~', so you would have to specify the full path</li>
 
<li>NOTE: the conf file can't interpret ${SS} environment variables or '~', so you would have to specify the full path</li>
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<li>We just used the command-line option for this tutorial since this path will vary by user.</li>
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<li>We just used the command-line option for this tutorial since this path will vary by user when running outside the workshop.</li>
 
</ul>
 
</ul>
 
</div>
 
</div>
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Now that we have all of our input files, we need just a simple command to run:
 
Now that we have all of our input files, we need just a simple command to run:
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  ${GC}/gotcloud snpcall --conf ${SS}/gotcloud.conf --numjobs 4 --region 22:36000000-37000000 --base_prefix ${SS} --outdir ${OUT}
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* When running at home if you don't have 6 CPUs, reduce the --numjobs setting (it will take longer to run).
 +
  ${GC}/gotcloud snpcall --conf ${SS}/gotcloud.conf --numjobs 6 --region 22:36000000-37000000 --base_prefix ${SS} --outdir ${OUT}
 
* <code>${GC}/gotcloud</code> runs GotCloud
 
* <code>${GC}/gotcloud</code> runs GotCloud
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* <code>align</code> tells GotCloud you want to run the alignment pipeline.
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* <code>snpcall</code> tells GotCloud you want to run the snpcall pipeline.
 
* <code>--conf</code> tells GotCloud the name of the configuration file to use.
 
* <code>--conf</code> tells GotCloud the name of the configuration file to use.
 
** The configuration for this test was downloaded with the seqshop input files.
 
** The configuration for this test was downloaded with the seqshop input files.
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** The Configuration file cannot read environment variables, so we need to tell GotCloud the path to the input files, ${SS}
 
** The Configuration file cannot read environment variables, so we need to tell GotCloud the path to the input files, ${SS}
 
** Alternatively, gotcloud.conf could be updated to specify the full paths
 
** Alternatively, gotcloud.conf could be updated to specify the full paths
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* <code>--out_dir</code> tells GotCloud where to write the output.
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* <code>--outdir</code> tells GotCloud where to write the output.
 
** This could be specified in gotcloud.conf, but to allow you to use the ${OUT} to change the output location, it is specified on the command-line
 
** This could be specified in gotcloud.conf, but to allow you to use the ${OUT} to change the output location, it is specified on the command-line
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</div>
 
</div>
 
</div>
 
</div>
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This should take about 5-7 minutes to run.
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This should take about 5-8 minutes to run.
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* After about 4 minutes of running, GotCloud snpcall will output some text to the screen.  Don't worry, that is expected and is just output from some of the intermediate tools.
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* It should end with a line like: <code>Commands finished in 402 secs with no errors reported</code>
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* It should end with a line like: <code>Commands finished in 329 secs with no errors reported</code>
      
If you cancelled GotCloud part way through, just rerun your GotCloud command and it will pick up where it left off.
 
If you cancelled GotCloud part way through, just rerun your GotCloud command and it will pick up where it left off.
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=== Running GotCloud Genotype Refinement ===
 
=== Running GotCloud Genotype Refinement ===
 
Since everything is setup, just run the following command (very similar to snpcall).
 
Since everything is setup, just run the following command (very similar to snpcall).
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  ${GC}/gotcloud ldrefine --conf ${SS}/gotcloud.conf --numjobs 2 --region 22:36000000-37000000 --base_prefix ${SS} --outdir ${OUT}
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  ${GC}/gotcloud ldrefine --conf ${SS}/gotcloud.conf --numjobs 6 --region 22:36000000-37000000 --base_prefix ${SS} --outdir ${OUT}
    
* Beagle will take about 1-3 minutes to complete
 
* Beagle will take about 1-3 minutes to complete
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Aren't you glad you didn't have to configure & run each one yourself?
 
Aren't you glad you didn't have to configure & run each one yourself?
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 +
 +
== Return to Workshop Wiki Page ==
 +
Return to main workshop wiki page: [[SeqShop: December 2014]]
Kleckner
87

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