Introduction

Warning

We are currently in the process of updating this tutorial for the new subtomogram averaging pipeline in relion-5.0. Please be patient while we work on this.

This tutorial will walk you through the tomography and sub-tomogram averaging pipeline of relion-5.0 . This semi-automated pipeline is designed to carry out all of the steps in a typical subtomogram averaging project from your raw tilt series movies to your final subtomogram average. We will use a test data set composed of 5 tomograms of immature HIV-1 dMACANC VLPs, which is available at EMPIAR-10164.

Requirements

  1. Your raw tilt series data as individual movies, or as individual motion-corrected images, but not as combined tilt series stacks.

  2. The mdoc files containing the metadata for each tilt series.

Getting Organised

We recommend to create a single directory per project. We’ll call this the project directory.

To start processing, make your own project directory and then download and unpack the raw data into this directory using the command below:

wget ftp://ftp.mrc-lmb.cam.ac.uk/pub/scheres/relion50_sta_tutorial_data.tar
tar -xf relion50_sta_tutorial_data.tar

We will start by launching the relion GUI. It is important to always launch the RELION graphical user-interface (GUI) from the project directory. To prevent errors with this, the GUI will ask for confirmation the first time you launch it in a new directory. Make sure you are inside the project directory, and launch the GUI by typing:

relion --tomo&

and answer Yes when prompted to set up a new relion project here.