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Shintaro Katayama edited this page Apr 13, 2016 · 40 revisions

Tutorials

How to run the pipeline on UPPMAX?

Installation

git clone -b v3dev https://github.com/shka/STRTprep.git STRTprep3.test
cd STRTprep3.test
bin/install_uppmax.sh

Protocol

  • Step 1: (Copy pre-build indexes, or) build indexes of reference genome and transcriptome, like as below. See also a help document.
sbatch -A bXXXXXXX -t 12:00:00 -p core -n 1 <<EOF
#!/usr/bin/env bash
. bin/setup_uppmax.sh
bin/index_hg19_ercc92_ynbA_u13369.sh
bin/index_refGene.sh hg19 src/ebwt/hg19_ercc92_ynbA_u13369/ref
EOF
  • Step 2: Edit conf.yaml and src/samples.csv
  • Step 3: Run preprocessing and quality-check as follows, then check out/byGene/samples.xls
    • N at (i) should be equal to total lanes of sequences but the maximum is 16.
    • Please put job number of (i) into Xi at (ii), and of (ii) into Xii at (iii); then the latter jobs will run if the former jobs succeeded.
    • ... Please contact when you process multiple libraries urgently - there is a method to run the pipeline more faster.
sbatch -A b2014069 -t 24:00:00 -p core -n N <<EOF ## (i)
#!/usr/bin/env bash
. bin/setup_uppmax.sh
PROCS=16 rake -m -j N qc1 qc2
EOF
sbatch -A b2014069 -t 24:00:00 -p core -n 2 -d afterok:Xi <<EOF ## (ii)
#!/usr/bin/env bash
. bin/setup_uppmax.sh
PROCS=16 rake -m -j 2 qc3
EOF
sbatch -A b2014069 -t 24:00:00 -p node -n 16 -d afterok:Xii <<EOF ## (iii)
#!/usr/bin/env bash
. bin/setup_uppmax.sh
PROCS=16 rake -m -j 16 qc
EOF
  • Step 4: Run differential expression tests and the down-stream analysis by plugins (for gene-based quantification) as follows.
    • Please extend time of job allocation (one hour), and change name of queue (devcore to core), when you met timeout
    • Please extend number of CPU allocation (one) when you met memory overflow
sbatch -A b2014069 -t 1:00:00 -p devcore -n 1 <<EOF
#!/usr/bin/env bash
. bin/setup_uppmax.sh
PROCS=16 rake plugins_gene
EOF
  • [v3dev] Step 5: Add "CLASS.TFE" column for classification of samples into src/samples.csv table, run differential expression tests, based on transcription-start-regions, then check out/byTFE/diffexp.xls and analyze more further!
    • Yi at (ii) is a job ID of (i).
sbatch -A b2014069 -t 24:00:00 -p core -n 8 <<EOF ## (i)
#!/usr/bin/env bash
. bin/setup_uppmax.sh
PROCS=16 rake tfe1
EOF
sbatch -A b2014069 -t 24:00:00 -p node -n 16 -d afterok:Yi <<EOF ## (ii)
#!/usr/bin/env bash
. bin/setup_uppmax.sh
PROCS=16 rake -m -j 16 tfe2
EOF
  • Step 6: Run differential expression tests and the down-stream analysis by plugins for TFE-based quantification as follows.
    • Please extend time of job allocation (one hour), and change name of queue (devcore to core), when you met timeout
    • Please extend number of CPU allocation (one) when you met memory overflow
sbatch -A b2014069 -t 1:00:00 -p devcore -n 1 <<EOF
#!/usr/bin/env bash
. bin/setup_uppmax.sh
PROCS=16 rake plugins_tfe
EOF

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