SRAToolKit-Sapelo2: Difference between revisions

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Sapelo2
Sapelo2
=== Version ===
=== Version ===
2.9.6-1,2.10.8,2.11.1
3.0.1, 3.0.3
===Author / Distributor===
===Author / Distributor===
Please see https://github.com/ncbi/sra-tools
Please see https://github.com/ncbi/sra-tools
===Description===
===Description===
The SRA Toolkit from NCBI is a collection of tools and libraries for using data in the INSDC Sequence Read Archives. The Sequence Read Archives (SRA) store raw sequence data from "next-generation" sequencing technologies including Illumina, 454, IonTorrent, Complete Genomics, PacBio and OxfordNanopores. In addition to raw sequence data, SRA now stores alignment information in the form of read placements on a reference sequence. The SRA-Toolkit includes the following tools:
The SRA Toolkit from NCBI is a collection of tools and libraries for using data in the INSDC Sequence Read Archives. The Sequence Read Archives (SRA) store raw sequence data from "next-generation" sequencing technologies including Illumina, 454, IonTorrent, Complete Genomics, PacBio and OxfordNanopores. In addition to raw sequence data, SRA now stores alignment information in the form of read placements on a reference sequence. The SRA-Toolkit includes the following tools:
{| class="wikitable"
|+
!Command
!Description
|-
|<code>fastq-dump</code>
|Convert SRA data into fastq format
|-
|<code>prefetch</code>
|Allows command-line downloading of SRA, dbGaP, and ADSP data
|-
|<code>sam-dump</code>
|Convert SRA data to sam format
|-
|<code>sra-pileup</code>
|Generate pileup statistics on aligned SRA data
|-
|<code>vdb-config</code>
|Display and modify VDB configuration information
|-
|<code>vdb-decrypt</code>
|Decrypt non-SRA dbGaP data ("phenotype data")
|-
|<code>abi-dump</code>
|Convert SRA data into ABI format (csfasta / qual)
|-
|<code>illumina-dump</code>
|Convert SRA data into Illumina native formats (qseq, etc.)
|-
|<code>sff-dump</code>
|Convert SRA data to sff format
|-
|<code>sra-stat</code>
|Generate statistics about SRA data (quality distribution, etc.)
|-
|<code>vdb-dump</code>
|Output the native VDB format of SRA data.
|-
|<code>vdb-encrypt</code>
|Encrypt non-SRA dbGaP data ("phenotype data")
|-
|<code>vdb-validate</code>
|Validate the integrity of downloaded SRA data
|}


fastq-dump: Convert SRA data into fastq format
=== Downloading SRA Data ===


prefetch: Allows command-line downloading of SRA, dbGaP, and ADSP data
The last version of this application is at /apps/eb/SRA-Toolkit/3.0.3-gompi-2022a


sam-dump: Convert SRA data to sam format
To use this version, please load the module with
<pre class="gscript">
ml SRA-Toolkit/3.0.3-gompi-2022a
</pre>


sra-pileup: Generate pileup statistics on aligned SRA data
You can download SRA data to local directory with the prefetch tool. This program downloads Runs (sequence files in the compressed SRA format) and all additional data necessary to convert the Run from the SRA format to a more commonly used format. You can search for a dataset using the search bar at the top of the SRA homepage. https://www.ncbi.nlm.nih.gov/sra Once you find a dataset you would like to download, search for the "Run number" in the table towards the bottom of the webpage for that dataset. Then create the folder where prefetch will deposit your files. This needs to be an empty folder. It is recommended that you download data into your scratch directory since there is plenty of space there.


vdb-config: Display and modify VDB configuration information
Next, run the command:


vdb-decrypt: Decrypt non-SRA dbGaP data ("phenotype data")
<pre>vdb-config --interactive</pre>


abi-dump: Convert SRA data into ABI format (csfasta / qual)
This will open a screen where you operate the buttons by pressing the letter highlighted in red, or by pressing the tab-key until the wanted button is reached and then pressing the space- or the enter-key. Make sure there is an X by the "'''Enable Remote Access'''" option on the MAIN tab, and X by the "'''enable local file-caching'''" option in the CACHE tab. Then set the "location of user-repository" to the empty folder you created. In the following image the data will be downloaded to /scratch/keekov/prefetchData. These settings are stored in a file called '''user-settings.mkfg''' in a hidden directory called .ncbi in the top level of your home directory.
[[File:Sratools.png|thumb]]


illumina-dump: Convert SRA data into Illumina native formats (qseq, etc.)
Then press "s" or navigate to the save button and press enter to save. Then press "x" or navigate to the exit button and press enter to exit. Now you can start the data download by running the command prefetch followed by the run number. For example the following downloads the dataset SRR390728.
<pre>prefetch SRR390728</pre>


sff-dump: Convert SRA data to sff format
For more information about the prefetch command refer to the [https://trace.ncbi.nlm.nih.gov/Traces/sra/sra.cgi?view=toolkit_doc&f=prefetch documentation]
 
sra-stat: Generate statistics about SRA data (quality distribution, etc.)
 
vdb-dump: Output the native VDB format of SRA data.
 
vdb-encrypt: Encrypt non-SRA dbGaP data ("phenotype data")
 
vdb-validate: Validate the integrity of downloaded SRA data


=== Running Program ===
=== Running Program ===
You can now run other SRA-Tools on the SRA data you've downloaded.
Also refer to [[Running Jobs on Sapelo2]]
Also refer to [[Running Jobs on Sapelo2]]


For more information on Environment Modules on Sapelo please see the [[Lmod]] page.
For more information on Environment Modules on Sapelo please see the [[Lmod]] page.


Here is an example of a shell script, sub.sh, to run on the batch queue:


<div class="gscript2">
<nowiki>#</nowiki>!/bin/bash<br>
<nowiki>#</nowiki>SBATCH --job-name=SRA-ToolsExample<br>
<nowiki>#</nowiki>SBATCH --partition=batch<br>       
<nowiki>#</nowiki>SBATCH --mail-type=ALL<br>
<nowiki>#</nowiki>SBATCH --mail-user=<u>username@uga.edu</u><br> 
<nowiki>#</nowiki>SBATCH --ntasks=<u>1</u><br> 
<nowiki>#</nowiki>SBATCH --mem=<u>10gb</u><br>   
<nowiki>#</nowiki>SBATCH --time=<u>08:00:00</u><br> 
<nowiki>#</nowiki>SBATCH --output=fastq-dump.%j.out<br>
<nowiki>#</nowiki>SBATCH --error=fastq-dump.%j.err<br>
cd $SLURM_SUBMIT_DIR<br>
ml SRA-Toolkit/3.0.3-gompi-2022a<br>   
fastq-dump -X 5 -Z /scratch/keekov/SRR390728.sra #You should use the directory you selected in the vdb-config step above<br> 
</div>


=== Documentation ===
=== Documentation ===

Latest revision as of 13:53, 25 June 2024

Category

BioInformatics

Program On

Sapelo2

Version

3.0.1, 3.0.3

Author / Distributor

Please see https://github.com/ncbi/sra-tools

Description

The SRA Toolkit from NCBI is a collection of tools and libraries for using data in the INSDC Sequence Read Archives. The Sequence Read Archives (SRA) store raw sequence data from "next-generation" sequencing technologies including Illumina, 454, IonTorrent, Complete Genomics, PacBio and OxfordNanopores. In addition to raw sequence data, SRA now stores alignment information in the form of read placements on a reference sequence. The SRA-Toolkit includes the following tools:

Command Description
fastq-dump Convert SRA data into fastq format
prefetch Allows command-line downloading of SRA, dbGaP, and ADSP data
sam-dump Convert SRA data to sam format
sra-pileup Generate pileup statistics on aligned SRA data
vdb-config Display and modify VDB configuration information
vdb-decrypt Decrypt non-SRA dbGaP data ("phenotype data")
abi-dump Convert SRA data into ABI format (csfasta / qual)
illumina-dump Convert SRA data into Illumina native formats (qseq, etc.)
sff-dump Convert SRA data to sff format
sra-stat Generate statistics about SRA data (quality distribution, etc.)
vdb-dump Output the native VDB format of SRA data.
vdb-encrypt Encrypt non-SRA dbGaP data ("phenotype data")
vdb-validate Validate the integrity of downloaded SRA data

Downloading SRA Data

The last version of this application is at /apps/eb/SRA-Toolkit/3.0.3-gompi-2022a

To use this version, please load the module with 

ml SRA-Toolkit/3.0.3-gompi-2022a

You can download SRA data to local directory with the prefetch tool. This program downloads Runs (sequence files in the compressed SRA format) and all additional data necessary to convert the Run from the SRA format to a more commonly used format. You can search for a dataset using the search bar at the top of the SRA homepage. https://www.ncbi.nlm.nih.gov/sra Once you find a dataset you would like to download, search for the "Run number" in the table towards the bottom of the webpage for that dataset. Then create the folder where prefetch will deposit your files. This needs to be an empty folder. It is recommended that you download data into your scratch directory since there is plenty of space there.

Next, run the command: 

vdb-config --interactive

This will open a screen where you operate the buttons by pressing the letter highlighted in red, or by pressing the tab-key until the wanted button is reached and then pressing the space- or the enter-key. Make sure there is an X by the "Enable Remote Access" option on the MAIN tab, and X by the "enable local file-caching" option in the CACHE tab. Then set the "location of user-repository" to the empty folder you created. In the following image the data will be downloaded to /scratch/keekov/prefetchData. These settings are stored in a file called user-settings.mkfg in a hidden directory called .ncbi in the top level of your home directory.

Sratools.png

Then press "s" or navigate to the save button and press enter to save. Then press "x" or navigate to the exit button and press enter to exit. Now you can start the data download by running the command prefetch followed by the run number. For example the following downloads the dataset SRR390728. 

prefetch SRR390728

For more information about the prefetch command refer to the documentation

Running Program

You can now run other SRA-Tools on the SRA data you've downloaded. Also refer to Running Jobs on Sapelo2

For more information on Environment Modules on Sapelo please see the Lmod page.

Here is an example of a shell script, sub.sh, to run on the batch queue:

#!/bin/bash
#SBATCH --job-name=SRA-ToolsExample
#SBATCH --partition=batch
#SBATCH --mail-type=ALL
#SBATCH --mail-user=username@uga.edu
#SBATCH --ntasks=1
#SBATCH --mem=10gb
#SBATCH --time=08:00:00
#SBATCH --output=fastq-dump.%j.out
#SBATCH --error=fastq-dump.%j.err

cd $SLURM_SUBMIT_DIR
ml SRA-Toolkit/3.0.3-gompi-2022a
fastq-dump -X 5 -Z /scratch/keekov/SRR390728.sra #You should use the directory you selected in the vdb-config step above

Documentation

Please see https://trace.ncbi.nlm.nih.gov/Traces/sra/sra.cgi?view=toolkit_doc for the documentation of each tool.

Installation

System

64-bit Linux

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