Welcome to the bulk RNAseq workshop

Required preparation

You are expected to make sure you can sign in to UCloud, SDU’s HPC platform on which we will be running this course. All data, assignments, and tools will be provided on UCloud. Please use your university ID to sign in (instructions below). If you run into problems, please write us (respond to the email that got you to this page).

Access Sandbox resources

Our first choice is to provide all the training materials, tutorials, and tools as interactive apps on UCloud, the supercomputer located at the University of Southern Denmark. Anyone using these resources needs the following:

  1. a Danish university ID so you can sign on to UCloud via WAYF1.

 

for UCloud Access click here

 

  1. basic ability to navigate in Linux/RStudio/Jupyter. You don’t need to be an expert, but it is beyond our ambitions (and course material) to teach you how to code from zero and how to run analyses simultaneously. We recommend a basic R or Python course before diving in.

  2. For workshop participants: Use our invite link to the correct UCloud workspace that will be shared on the day of the workshop. This way, we can provide you with compute resources for the active sessions of the workshop2 Click the link below after your first uCloud access and accept the invite that shows.

 

Invite link to uCloud workspace

 

  1. Additional files needed for exercises from Day 1 - Lecture 2 (QC report files)

  1. Course slides

 

  1. COURSE EVALUATION SURVEY - The Novo Nordisk Foundation funds the Sandbox project and is interested in the outcomes of our training activities, so we really appreciate your responses!

 

Evaluation survey link

 

Workshop Agenda

Day 1

Time Activity
9:00 Intro to Course
9:15 Experimental Design
9:45 Preprocessing and Library Prep
10:15 Coffee break
10:30 Trimming, QC, and Alignment
11:30 Feature counts and MultiQC
12:00 Lunch break
13:00 Feature counts, MultiQC, and Alignment
13:45 Intro to HPC and UCloud
14:30 Coffee break
15:00 Nextflow pipelines, nf-core, and UCloud
16:00 RNAseq results from pipeline
16:30 Q & A

Day 2

Time Activity
9:00 UCloud setup / recap
9:30 RNAseq count matrix and normalization
10:15 Coffee break
10:30 Exercise: Count matrices
11:30 Exploratory data analysis
12:00 Lunch break
13:00 Exercise: Exploratory data analysis
14:30 Coffee break
14:45 Differential Expression Analysis
16:30 Q & A

Day 3

Time Activity
9:00 UCloud setup / recap
9:45 DEA visualization
10:30 Coffee break
10:45 Gene annotation and databases
11:15 Exercise: Gene annotation
12:00 Lunch break
13:00 Gene annotation exercise
13:30 Functional analysis
14:15 Coffee break
14:30 Exercise: Functional analysis
15:15 Workflow summary
15:30 Bring your own data
16:30 Wrap-up & course eval

Workshop

The Health Data Science Sandbox aims to be a training resource for bioinformaticians, data scientists, and those generally curious about how to investigate large biomedical datasets. We are an active and developing project seeking interested users (both trainees and educators). All of our open-source materials are available on our Github page and can be used on a computing cluster! We work with both UCloud, GenomeDK and Computerome, the major Danish academic supercomputers.

Transcriptomics apps

High-Performance Computing (HPC) platforms are essential for large-scale data analysis. Therefore, we will run our bulk RNA-seq analyses on one of the national HPC platforms, UCloud.

  • If you want to review the course material, visit our website where you will find the content for all the lectures.
  • Zenodo link to download the material (slides, assignments, data, etc.) for this workshop here.
  • To get started with our transcriptomics app, follow the UCloud setup guidelines. This will help you set up a new job and repeat the exercises on your own.
  • To run the nf-core RNAseq pipeline follow the instructions here. This will generate the output from the preprocessing pipeline.
Image

Transcriptomics Sandbox: Our sandbox for bulk or single-cell RNA sequencing analysis provides stand-alone analysis and visualization tools.

 

We are developing other apps. If you are interested, explore the modules section on our website!

Discussion and feedback

We hope you enjoyed the workshop. If you have broader questions, suggestions, or concerns, now is the time to raise them! Remember that you can check out longer versions of our tutorials as well as other topics and tools in each of the Sandbox modules. We regularly run workshops on a variety of health data science topics that you can also check out (follow our news here).

As data scientists, we also would be happy for some quantifiable info and feedback - we want to build things that the Danish health data science community is excited to use.

 

Nice meeting you and we hope to see you again!

Footnotes

  1. Other institutions (e.g. hospitals, libraries, …) can log on through WAYF. See all institutions here↩︎

  2. To use Sandbox materials outside of the workshop: remember that each new user has hundreds of hours of free computing credit and around 50GB of free storage, which can be used to run any uCloud software. If you run out of credit (which takes a long time) you’ll need to check with the local DeiC office at your university about how to request compute hours on UCloud. Contact us at the Sandbox if you need help or want more information.↩︎