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Forum: How people usually perform scRNA Seq Analysis on HPC?

I am beginner and want to know how people usually perform single cell RNA Seq Analysis on clusters - RStudio GUI on HPC or RScript execution on HPC terminal.

I heard that one way to do is - write script in a section, execute, check errors, execute and then finally compile all together to execute final one. Same can be applied for other samples or cells. Also, is this the way to develop a new pipeline?

Any comments would be appreciated.

hpc single-cell cluster scrna-seq
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updated by Ram Ram • written by Tundup • Tundup

If you have a HPC then you must have folks that administer that HPC. You can work with them to see if any of this is already available on your cluster (there may already be RStudio etc). You could then ask them to install the necessary packages (if they are not already present, you won't likely be the only person using scRNA at your institution). This may lead to a much more efficient/pleasant user experience.

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written by GenoMax GenoMax

3 answers

You can send a job in your HPC to set up a Rstudio instance or Jupyterlab instance inside containers environments like Docker or Singularity. Once the instance is running you will get the URL of your instance in the log file of the job. Then you can make a tunneling (see below) between your HPC and your workstation and paste the instance URL inside your favorite web browser.

ssh -t -t name@hpc.dot -L port:localhost:port ssh node -L port:localhost:port

Everything you will do in this Rstudio instance will be processed by the HPC with the specification you asked in the job.

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written by Bastien Hervé Bastien Hervé

This assumes that the HPC is setup to allow port-forwarding etc. Many HPC may be tightly controlled. The speed of internet connection/firewalls/VPN can further constrain the entire user experience of this method. Something to keep in mind.

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written by GenoMax GenoMax

That is why I prefer Singularity over Docker at the moment, sudo privileges are not required to run a singularity instance but docker does.

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written by Bastien Hervé Bastien Hervé

Your HPC cluster may provide an instance of Open OnDemand that allows you to access the cluster and run various applications like Rstudio directly from your browser.

Because every scRNA-seq experiment is so different, I find it hard to write a one-size-fits-all script or pipeline to do the analysis. Instead, I end up doing a lot of experimentation and iteration in Rstudio. If your cluster admins provides Open OnDemand for running Rstudio, then I would recommend using it there. If not, I would recommend asking them about it. It's not really a lot of work to setup, and it's extremely useful.

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written by Dave Carlson Dave Carlson
  1. Set up an R Environment & install essential packages on HPC;
  2. Write scripts and do some tests in local RStudio (excluding mem-intensed parts);
  3. Run the whole pipeline on HPC, simply Rscript --vanilla my_script.R

BTW sometimes the first step can be the most difficult :)

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written by gglim • gglim

I would strongly recommend a containerized solution to make things software-wise reproducible and portable.

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written by ATpoint ATpoint

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