Linux & Shell Scripting Small Group Lecture 3 How to Learn to Code Workshop group/ Erin Osborne Nishimura 1

Where are we? SessionDateTopicSlidesExercises 1June 15, 1pmLet's do some linux 2June 22, 1pmLet's do some linux 3TUESDAY June 30, 1pm Let's execute jobs on the cluster 4July 6, 1pm Automating workflows 5July 13, 1pmBuilding shell scripts 6July 20, 1pm Shell scripts with bells and whistles 7TBA Collaborating with others; Documenting our work 8Aug 3, pm Future directions for users and the course

Questions & Comments

Group pop quiz What does this do? $ ls 1_cdc1.txt 2_cdc2.txt 3_cdc3.txt 4_cdc4.txt 5_cdc5.txt my_cyclins_README.log my_cyclins_downloaddata.txt $ wc *.txt | sort > captured.txt How would you unpack this? project_update.tar.gz You have a file that contains a list of Arabidopsis transcription factor names: "ID","Name","Species","GeneID","Family","Evidence" "T000676_1.01","ANT","Arabidopsis_thaliana","AT4G37750-TAIR-G","AP2","D" "T000588_1.01","AT1G22985","Arabidopsis_thaliana","AT1G22985-TAIR-G","AP2","D" "T000614_1.01","AT1G75490","Arabidopsis_thaliana","AT1G75490-TAIR-G","AP2","D" What all the IDs of transcription factors that contain the bHLH domain? How many are there?

Learning objectives week 2 Move files to and from kure Decompress and unpackage a tarball Read files Start to use the wild card * Start to chain commands together using |

Week 2 Killdevil module Bsub

No seriously, what is Killdevil? 7

Killdevil is a high-performance computing environment Linux operating system 1 login node 774 compute nodes – 48 – 96 GB memory per node. – 12 – 16 CPU’s cores per node. 2 large memory nodes (1 TB) 12 Graphics Processors (GPUs) nodes File systems for storage 8

If a job takes more than 3 seconds to complete, cancel it! Use bsub instead!

Load File Sharing (LSF) LSF -- allocates nodes to job submissions -- max 64 on kure, 1024 on killdevil -- fair scheduling -- takes into consideration your status, queue, your job, recent activity, node requirements $ bqueues #This will show you all the ‘queues’ $ bjobs -u all #This will show all the jobs right now!

Execute big jobs on kure using bsub Execute jobs on kure with bsub to get off the head node $ bsub -q week [-n 1] [ -M ] [-o %J.log] “ ” Use the queue called ‘week’; I want one node; Give me more memory! output anything about how this job runs to a log file. $ bjobs# check all running jobs $ bpeek [NUMBER]# see the screen output of a job $ bkill # terminate a job

Big jobs – exercise #1 The directory /proj/seq/data/ contains lots of whole genome sequencing resources. These are big files. – Go to /proj/seq/data/ Now navigate to the directory: – /proj/seq/data/ce10_NCBI/Sequence/WholeGenomeFasta – This directory contains the C. elegans genome (genome.fa). Try to count the lines in this genome. If it takes longer than 5 seconds, cancel out. Next, re-submit the command using bsub: /proj/seq/data/ce10_NCBI/Sequence/WholeGenomeFasta/genome.fa $ bsub -q week “wc genome.fa” Check your job with “bpeek” and “bjobs”. When your job is finished, check your . Try to save the output of your job to a logfile in your home directory: $ bsub –q week –o ~/bjobs_output_%J.log “wc genome.fa” Watch your job with “bpeek” and “bjobs”. Check your . Anything there? Go to your home directory. Anything there?

Parallel processing $ bsub -q week -n -R “span[hosts=1]” [ -M ] [-o %J.log] “ ” You must use –n and –R together when submitting parallel jobs -n #This is the number of nodes (also called jobs) to use. Maximum of 12. -R “span[hosts=1]” #This tells LSF to put all of those nodes/jobs on one host.

Modules on killdevil $ module avail – View available modules on killdevil $ module list -List the modules you have loaded up and ready to use $ module add $ module load – These are the same. They both load an available module into your list of usable modules. They will load for your ssh session and be removed when you log out. $ module initadd – Load this module every time I log in, as soon as I log in $ module initrm – Remove this module from the list of all modules that load as soon as I log in. $ module $ module –H $ module –help – These are the same. They all take you to the help page

Exercise 2 Browse the list of available modules Do you recognize any of these applications? Which have you used before? Load the module bedtools to your list of available modules Read a little bit about bedtools using its manual page Read about bedtools getfasta What does bedtools getfasta do? Load another module, maybe something you recognize. Can you find the manual or help message for this utility? (Hint, the point of this is that some of these are really hard to find manuals for and that you need to go to the internet for help; Sometimes that doesn’t even work).

An intro to Exercise 3 Fasta files – Genome sequences are kept as fasta files. – Bed files – Gives the intervals of a genome – Gff files – Gives intervals of a genome –

Exercise 3A Let’s use bedtools getfasta. The point of this exercise will be to make a fasta file containing the sequences of all the genes that are located on the left hand of chromosome I in the yeast genome. Start a README.txt file for this project Obtain a gtf/gff file of the entire yeast genome. This is the full annotation of every gene in the yeast genome – Go to UCSC genome browser’s table page: – – Download a gtf file by filling out the form to exactly match the next slide…

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Exercise 3B (cont) Download the.gtf.gz file to your local computer Upload it to killdevil. Unzip it using $ gunzip Look inside of it to see what a.gtf file looks like. Compare the file with the description of a gtf file on UCSC genome browser (slide 16)

Exercise3C Make a smaller annotation file that contains just the coding sequences (“CDS”) of genes that are on the left side of chromosome 1. These genes will all start with the letters “YAL”. – Filter for “YAL”; Filter for “CDS”; save the results in the file chrI_left_CDS.gtf – How many genes are in there? Execute bedtools. Learn how to use bedtools. – Input – The yeast genome. We downloaded this in week2; exercise2. I think it’s called S_cerevisiae.fa. You will need to know the FULL PATH to this file. – Input – The gtf file called chrI_left_CDS.gtf – Input – The NAME you want to give to your output fasta file. Use the bedtools getfasta help pages to figure out how to use this code. – Hint: you will need the arguments (-fi, -bed, and -fo, and one other option) Check that all your sequences start with an “ATG”. Check that you have the correct number of fasta sequence entries (“>”).

Exercise 3D Complete your README.txt file