Selected article for: "Îl reaction and RNA template"
Author: Wanchao Yin; Chunyou Mao; Xiaodong Luan; Dan-Dan Shen; Qingya Shen; Haixia Su; Xiaoxi Wang; Fulai Zhou; Wenfeng Zhao; Minqi Gao; Shenghai Chang; Yuan-Chao Xie; Guanghui Tian; He-Wei Jiang; Sheng-Ce Tao; Jingshan Shen; Yi Jiang; Hualiang Jiang; Yechun Xu; Shuyang Zhang; Yan Zhang; H. Eric Xu
Title: Structural Basis for the Inhibition of the RNA-Dependent RNA Polymerase from SARS-CoV-2 by Remdesivir
  • Document date: 2020_4_9
    • ID: 7v7pzclb_35
    Snippet: A gel mobility shift assay was performed to detect RNA binding by the RdRp complex. The binding reaction contained 25 mM HEPES pH 7.4, 100 mM sodium chloride, 2 mM magnesium chloride and 1 mM TCEP, 9 μg RdRp complex protein with increasing amounts of templateprimer RNA (0, 0.3, 0.6, 1.2 and 2 μg). For the nsp12 protein, the binding reaction was combined with 1.5 μg template-primer RNA. Binding reactions were incubated for 30 min at room temper.....
    Document: A gel mobility shift assay was performed to detect RNA binding by the RdRp complex. The binding reaction contained 25 mM HEPES pH 7.4, 100 mM sodium chloride, 2 mM magnesium chloride and 1 mM TCEP, 9 μg RdRp complex protein with increasing amounts of templateprimer RNA (0, 0.3, 0.6, 1.2 and 2 μg). For the nsp12 protein, the binding reaction was combined with 1.5 μg template-primer RNA. Binding reactions were incubated for 30 min at room temperature and resolved on 4-20% native polyacrylamide gel (Thermo Fisher Scientific) running in 1×TBE buffer at 90 V for 1h in 4 °C cool-room. Then the gel was carefully taken out and was dyed with Ultra GelRed Nucleic Acid Stain (Vazyme Biotech Co.,Ltd) according to the manufacturer's protocol. The dye-stsined gel was visualized on a BIO-RAD Fluorescence Imager.

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