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Biochemistry

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JoVE Journal Biochemistry
Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon
 

Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon

Article DOI: 10.3791/64136-v • 04:52 min • July 1st, 2022
July 1st, 2022 •
1, 1, 2, 2,3, 1,4,5
1Laboratory of Genetics, The Salk Institute for Biological Studies, 2Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, 3Department of Chemistry and Biochemistry, University of California San Diego, 4Graduate School of Biological Sciences, Section of Molecular Biology, University of California San Diego, 5Department of Integrative Structural and Computational Biology, The Scripps Research Institute

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The present protocol describes a generalized and easy-to-implement scheme for tilted single-particle data collection in cryo-EM experiments. Such a procedure is especially useful for obtaining a high-quality EM map for samples suffering from preferential orientation bias due to adherence to the air-water interface.

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Keywords: Cryo-EM Single-particle Analysis Stage Tilt Particle Orientation Data Collection Eucentric Height Image Shift Focus Hole Finding Automated Targeting
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