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General and robust covalently linked graphene oxide affinity grids for high-resolution cryo-EM

By Feng Wang, Yanxin Liu, Zanlin Yu, Sam Li, Yifan Cheng, David A. Agard

Posted 02 Jun 2019
bioRxiv DOI: 10.1101/657411 (published DOI: 10.1073/pnas.2009707117)

Despite their great potential to facilitate rapid preparation of quite impure samples, affinity grids have not yet been widely employed in single particle cryo-EM. Here, we chemically functionalize graphene oxide coated grids and use a highly specific covalent affinity tag system. Importantly, our polyethylene glycol spacer keeps particles away from the air-water interface and graphene oxide surface, protecting them from denaturation or aggregation and permits high-resolution reconstructions of small particles.

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