Selected article for: "Ã pixel size and electron microscope"
Author: Yan Gao; Liming Yan; Yucen Huang; Fengjiang Liu; Yao Zhao; Lin Cao; Tao Wang; Qianqian Sun; Zhenhua Ming; Lianqi Zhang; Ji Ge; Litao Zheng; Ying Zhang; Haofeng Wang; Yan Zhu; Chen Zhu; Tianyu Hu; Tian Hua; Bing Zhang; Xiuna Yang; Jun Li; Haitao Yang; Zhijie Liu; Wenqing Xu; Luke W. Guddat; Quan Wang; Zhiyong Lou; Zihe Rao
Title: Structure of RNA-dependent RNA polymerase from 2019-nCoV, a major antiviral drug target
  • Document date: 2020_3_17
    • ID: glfxrla9_22
    Snippet: Cryo-EM sample preparation and data collection 25 An aliquot of 3 μL of the sample at 0.7 mg/mL (added with 0.025% DDM) was applied onto a H2/O2 glow-discharged, 300-mesh Quantifoil R1.2/1.3 grid (Quantifoil, Micro Tools GmbH, Germany). The grid was then blotted for 3.0 s with a blot force of 0 at 8°C and 100% humidity and plunge-frozen in liquid ethane using a Vitrobot (Thermo Fisher Scientific, USA). Cryo-EM data were collected with a 300 keV.....
    Document: Cryo-EM sample preparation and data collection 25 An aliquot of 3 μL of the sample at 0.7 mg/mL (added with 0.025% DDM) was applied onto a H2/O2 glow-discharged, 300-mesh Quantifoil R1.2/1.3 grid (Quantifoil, Micro Tools GmbH, Germany). The grid was then blotted for 3.0 s with a blot force of 0 at 8°C and 100% humidity and plunge-frozen in liquid ethane using a Vitrobot (Thermo Fisher Scientific, USA). Cryo-EM data were collected with a 300 keV Titan Krios electron microscope (Thermo Fisher Scientific, 30 USA) and a K2 Summit direct electron detector (Gatan, USA). Images were recorded at EFTEM with a 165000× magnification and calibrated super-resolution pixel size 0.82 Å/pixel. The exposure time was set to 5 s with a total accumulated dose of 60 electrons per Å 2 . All images were automatically recorded using SerialEM (21). For Dataset-1, a total of 7,994 images were collected with a defocus range from 1.0 μm to 1.8 μm. For Dataset-2 (under reducing conditions), a total of 35 8494 images were collected with a defocus range from 1.1 μm to 2.0 μm. Statistics for data collection and refinement are in Table S1 .

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