Rneasy96 extraction kit

Example 6

Anti-Norovirus Activity

Norwalk virus replicon assays were performed as reported by Constantini et al. (Antivir Ther 2012, 17, 981-991). HG23 cells (derived from Huh-7 cells) containing NoV replicon RNA are seeded at a density of 3,000 cells/well in 96-well plates and incubated at 37° C. and 5% CO₂ overnight. Compounds were tested at concentrations ranging from 0.1 to 100 μM. Compounds were added in triplicate to 80 to 90% confluent monolayers and incubated at 37° C. and 5% CO₂. Untreated cells were included in each plate. Following five days incubation (37° C., 5% CO₂), total cellular RNA was isolated with RNeasy96 extraction kit from Qiagen. Replicon RNA and an internal control (TaqMan rRNA control reagents, Applied Biosystems) were amplified in a single step.

The median effective concentrations (EC₅₀) ranges of several of the compounds described herein against NoV are shown in Table 3:

Example 6

Anti-Norovirus Activity

Norwalk virus replicon assays were performed as reported by Constantini et al. (Antivir Ther 2012, 17, 981-991). HG23 cells (derived from Huh-7 cells) containing NoV replicon RNA are seeded at a density of 3,000 cells/well in 96-well plates and incubated at 37° C. and 5% CO₂ overnight. Compounds were tested at concentrations ranging from 0.1 to 100 μM. Compounds were added in triplicate to 80 to 90% confluent monolayers and incubated at 37° C. and 5% CO₂. Untreated cells were included in each plate. Following five days incubation (37° C., 5% CO₂), total cellular RNA was isolated with RNeasy96 extraction kit from Qiagen. Replicon RNA and an internal control (TaqMan rRNA control reagents, Applied Biosystems) were amplified in a single step.

The median effective concentrations (EC₅₀) ranges of several of the compounds described herein against NoV are shown in Table 3: