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First, we'll run breseq on a small data set to be sure that it is installed correctly, and to get a taste for what the output looks like. This sample is a mixed population of bacteriophage lambda that was co-evolved in lab with its E. coli hosts.

Data

The data files for this example are in the path:

Code Block
$BI/ngs_course/lambda_mixed_pop/data

Copy this directory to a new directory called BDIB_breseq in your $SCRATCH space and cd into it.

Code Block
languagebash
titleClick here for the solution
collapsetrue
cds
mkdir BDIB_breseq_lambda
cp $BI/ngs_course/lambda_mixed_pop/data/* BDIB_breseq_lambda
cd BDIB_breseq_lambda
ls 

If the copy worked correctly you should see  the following 2 files:

File Name

Description

Sample

lambda_mixed_population.fastq

Single-end Illumina 36-bp reads

Evolved lambda bacteriophage mixed population genome sequencing

lambda.gbk

Reference Genome

Bacteriophage lambda

Running breseq

Because this data set is relatively small (roughly 100x coverage of a 48,000 bp genome), a breseq run will take < 5 minutes. Submit this command to the TACC development queue or run on an idev node.

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languagebash
titlebreseq prep and commands

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Environment

To set your profile up to run breseq, we need to add "module load bowtie/2.1.0" to your profile.

Code Block
languagebash
titleAdding bowtie to your profile
cdh  #move to your home directory
echo "module load bowtie/2.1.0" >> .profile  #this command updates your profile to automatically load the bowtie module

After you've completed these commands, exit lonestar and re log in to re run your profile.  

Data

The data files for this example are in the path:

Code Block
$BI/ngs_course/lambda_mixed_pop/data

Copy this directory to a new directory called BDIB_breseq in your $SCRATCH space and cd into it.

Code Block
languagebash
titleClick here for the solution
collapsetrue
cds
mkdir BDIB_breseq_lambda
cp $BI/ngs_course/lambda_mixed_pop/data/* BDIB_breseq_lambda
cd BDIB_breseq_lambda
ls 

If the copy worked correctly you should see  the following 2 files:

File Name

Description

Sample

lambda_mixed_population.fastq

Single-end Illumina 36-bp reads

Evolved lambda bacteriophage mixed population genome sequencing

lambda.gbk

Reference Genome

Bacteriophage lambda

Running breseq

Because this data set is relatively small (roughly 100x coverage of a 48,000 bp genome), a breseq run will take < 5 minutes. Submit this command to the TACC development queue or run on an idev node.

Code Block
languagebash
titlebreseq prep and commands
idev  #idev starts an "interactive development" mode which allows you to run computationally intensive tasks
 
breseq -j 12 -r lambda.gbk lambda_mixed_population.fastq > log.txt

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Expand
titleClick here for commands file example and launcher_creator.py generator
Code Block
titleExample commands file
breseq -j 12 -r NC_012967.1.gbk -o output_00K SRR030252_1.fastq SRR030252_2.fastq &> 00K.log.txt
breseq -j 12 -r NC_012967.1.gbk -o output_02K SRR030253_1.fastq SRR030253_2.fastq &> 02K.log.txt
breseq -j 12 -r NC_012967.1.gbk -o output_05K SRR030254_1.fastq SRR030254_2.fastq &> 05K.log.txt
breseq -j 12 -r NC_012967.1.gbk -o output_10K SRR030255_1.fastq SRR030255_2.fastq &> 10K.log.txt
breseq -j 12 -r NC_012967.1.gbk -o output_15K SRR030256_1.fastq SRR030256_2.fastq &> 15K.log.txt
breseq -j 12 -r NC_012967.1.gbk -o output_20K SRR030257_1.fastq SRR030257_2.fastq &> 20K.log.txt
breseq -j 12 -r NC_012967.1.gbk -o output_40K SRR030258_1.fastq SRR030258_2.fastq &> 40K.log.txt
Code Block
launcher_creator.py -q normal -t 3:00:00 -n launcher -a "UT-2015-05-18" -m "module load bowtie/2.1.0"
qsub launcher.sge

This will likely sit for some time in the launcher que, making it a good opportunity to work through the interrogating launcher queue portion of our linux tutorial if you didn't get the opportunity to earlier.

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