| Assay Method Information | |
| | Rhinovirus Polymerase (HRV1bpol) and HCV Polymerase (HCVpol) Assays |
| Description: | The enzyme activity of hepatitis C virus RNA polymerase (HCVpol) and human rhinovirus 16 RNA polymerase (HRV16pol) is measured as an incorporation of tritiated NMP into acid-insoluble RNA products. HCVpol and HRV16pol assay reactions contain 30-100 nM recombinant enzyme, 50-500 nM heteropolymeric RNA, 0.5 μCi tritiated NTP, 0.1-1 μM of other NTPs, in a standard reaction buffer containing MgCl2. Enzymatic reactions are incubated for 2.5 hours at 30° C., in the presence of increasing concentration of inhibitor. At the end of the reaction, the total RNA is precipitated with 10% TCA, and acid-insoluble RNA products are filtered on a size exclusion 96-well plate. After washing of the plate, scintillation liquid is added and radiolabeled RNA products are detected according to standard procedures with a Trilux Microbeta scintillation counter. |
| Affinity data for this assay | |
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