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Charles University in Prague, Univerzita Karlova v Praze
Type Public, Ancient
Established - 1348; 673 years ago
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Structure of the SARS-CoV-2 RNA-dependent RNA polymerase in the presence of favipiravir-RTP
RNA Synthesis The RNA template, 5′-rUrUrUrUrUrCrArUrArArCrUrUrArArUrCrUrCrAr- CrArUrArGrCrArCrUrG-3′, and RNA primer 5′-rCrArGrUrGrCrUrArUrGrUr- GrArGrArUrUrArArGrUrUrArU-3′ were prepared by solid-phase synthesis on an ÄKTA oligopilot plus 10 (Cytiva). RNAs were cleaved from the solid support by treating with 4 mL of a 1:1 mixture of 28% wt NH3/H2O solution and 33% wt CH3NH2/EtOH solution at 55 °C for 30 min; then the silyl protecting groups were removed by treating with 3 mL of 1:1 mixture of triethylamine trihydrofluoride and dimethyl sulfoxide at 55 °C for 90 min. Then, 30 mL of cold 50 mM NaClO4 in acetone was added to precipitate the RNA product. After centrifugation, the pellet of RNA was dissolved in 5 mL of water and passed through a Sep-Pack C18 Cartridge, 5 g sorbent (Waters). Eluates containing RNA were combined and lyophilized. Mass spectroscopy of the template RNA found m/z ([M-7H+]) = 1,341.8 (theoretical 1,342.1) and found m/z ([M-6H+]) = 1,565.6 (theoretical 1,565.9); mass spectrometry of the primer RNA found m/z ([M-6H+]) = 1,278.6 (theoretical 1,278.9) and found m/z ([M-5H+]) = 1,534.5 (theoretical 1,534.9).
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I get nothing but things telling me about the Charles University in Prague. So would you be so kind as to provide what you are talking about? Thanks in advance.