Figure 2.
Effects of washing and reaction temperature. A. In vitro oxidized or non-oxidized radiolabeled RNA (1.8 kb) with or without derivatization, was incubated with strep-beads at 37°C for 15min. The beads were washed with several buffer solutions including 4M urea, and then heat denatured to dissociate the bound RNA. Radioactivity in the pull-down fraction as well as initial input RNA was quantified using a PhosphorImager. B. In vitro oxidized radiolabeled ARP-RNA was incubated with strep-beads at different temperatures for 30 min. The beads were washed several times with salt buffers including 4M urea buffer solution and heat denatured. Radioactivity in the pull-down fraction and initial input RNA was determined. The results are represented as means ± SD from triplicate samples.