Selected article for: "detailed procedure and RNA template"
Author: Jingyue Ju; Xiaoxu Li; Shiv Kumar; Steffen Jockusch; Minchen Chien; Chuanjuan Tao; Irina Morozova; Sergey Kalachikov; Robert N. Kirchdoerfer; James J. Russo
Title: Nucleotide Analogues as Inhibitors of SARS-CoV Polymerase
  • Document date: 2020_3_14
    • ID: hj675z1b_13
    Snippet: The sequence of the primer and template used for these extension reactions, which are within the N1 coding sequence of the SARS-CoV-2 genome, is shown at the top of the figure. Polymerase extension reactions were performed by incubating (a) 2'-F,Me-UTP, (b) 3'-F-dTTP, and (c) 3'-N 3 -dTTP with pre-assembled SARS-CoV polymerase (nsp12, nsp7 and nsp8), the indicated RNA template and primer, and the appropriate reaction buffer, followed by detection.....
    Document: The sequence of the primer and template used for these extension reactions, which are within the N1 coding sequence of the SARS-CoV-2 genome, is shown at the top of the figure. Polymerase extension reactions were performed by incubating (a) 2'-F,Me-UTP, (b) 3'-F-dTTP, and (c) 3'-N 3 -dTTP with pre-assembled SARS-CoV polymerase (nsp12, nsp7 and nsp8), the indicated RNA template and primer, and the appropriate reaction buffer, followed by detection of reaction products by MALDI-TOF MS. The detailed procedure is shown in the Methods section. For comparison, data for extension with UTP are presented in Extended Data Fig. 2 . The accuracy for m/z determination is ± 10 Da.

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