Figure 4.

Monitoring the synthesis of two different RNA transcripts in the same tube. Each reaction contained bacteriophage T7 RNA polymerase, fluorescein-labeled molecular beacon MB-poly(A)-FAM-2′-O-methyl and tetrachlorofluorescein-labeled molecular beacon MB-sinR-TET-2′-O-methyl. T7-rpoB-poly(A) DNA template was added to the first reaction (left), T7-sinR DNA template was added to the second reaction (middle) and both DNA templates were added to the third reaction (right). The fluorescence intensities of fluorescein (green lines) and tetrachlorofluorescein (red lines) were monitored simultaneously during each reaction. The results show that synthesis of rpoB poly(A) transcripts only elicits fluorescence from poly(A)-specific molecular beacons and synthesis of sinR transcripts only elicits fluorescence from sinR-specific molecular beacons.