Preparing frozen-hydrated protein–nucleic acid assemblies for high-resolution cryo-EM imaging

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Abstract

High-resolution image acquisition and structure determination by cryo-electron microscopy is becoming increasingly streamlined. Preparing electron-microscopy grids of suitable quality remains, however, a critical bottleneck. Strategies to achieve particle monodispersity, optimal sample concentration and suitable ice thickness can vary from specimen to specimen. In this book chapter we describe our protocols for negative-stain grid and cryo-grid preparation, which we apply to studying protein–nucleic acid complexes.

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Goswami, P., Locke, J., & Costa, A. (2018). Preparing frozen-hydrated protein–nucleic acid assemblies for high-resolution cryo-EM imaging. In Methods in Molecular Biology (Vol. 1814, pp. 287–296). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8591-3_17

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