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Cryo-tomography and subtomogram averaging are increasingly popular techniques for structural determination of macromolecular complexes in situ. They have the potential to achieve high-resolution views of native complexes, together with the details of their location relative to interacting molecules.

Tilt series of vitrified COPII-coated tubules are collected in a high-end cryo-EM.

Tomograms are then reconstructed by back-projection. We use SerialEM for data collection and IMOD for reconstructions.

tomo_pipeline.jpg

Repeating units within tomograms are averages through sub-tomogram averaging pipelines. We use a combination of home made scripts, scripts shared by the Briggs lab, and Dynamo, to process our data.

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