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. Author manuscript; available in PMC: 2013 Feb 13.
Published in final edited form as: Methods Enzymol. 2012;511:463–483. doi: 10.1016/B978-0-12-396546-2.00021-8

Figure 21.4.

Figure 21.4

An RNA-based HCV helicase assay suitable for HTS. (A) Design of the split beacon helicase assay (SBHA). In the RNA substrate, the bottom strand and the Cy5-labeled strand are RNA, while the IAbRQ-labeled strand is composed of DNA. In the DNA-labeled substrate, all three strands are composed of DNA, with Ts replacing Us in the bottom strand and the Cy5-labeled strand. (B) Time courses for sample negative control reactions using the DNA (closed circles) and the RNA (open circles) substrate, positive control reactions containing DNA plus 50 µM primuline (+), and the RNA substrate with 50 µM primuline (*). Lines show fit to the equations described in the text. The fluorescence observed in a well without substrate (×) is shown for comparison. (C) Reaction extent for two 96-well plates, one of which contained an RNA substrate and one a DNA substrate.