FIGURE 1.

Target RNA conversion into a primer for RCA, the impact of E. coli RNase III. The reactions were carried out under the conditions described in Materials and Methods. The 5′-end-labeled (A) RNA2-PP1 (panel 1), (B) RNA2-PP2 (panel 1), and RNA1-PP1 (panel 2) hybrids were incubated with Phi29 DNA polymerase in the absence or presence of dNTP. (A, panel 2) The experiments with E. coli RNase III were performed using RNA2-PP1 duplex. The RNA degradation and RCA products were analyzed by electrophoresis through denaturing 8% polyacrylamide gels. Different RNA-PP duplexes are shown above the gels. The radioactive label is indicated by the star. The contents of samples are shown above the gel lines. Reaction products are labeled as “a” for the 5′-end-labeled RNA1 hydrolysis product serving as an RNA primer for RCA and “b” for the 5′-end-labeled RCA product. (C) The reaction scheme represents the conversions of target RNA.