Figure 3.

Cotranscriptional assembly of Split-Broccoli with transcription terminator structures improves signal relative to independently transcribed and assembled RNA. When cotranscribed with equimolar amounts of template DNA in a one-pot in vitro transcription reaction designed to maximize time resolution, Split-Broccoli (Top-T + Bottom-T, green) exhibits approximately 88% of the signal generated by the unimolecular, unsplit variant (3WJdB-T, black) after a 4-h reaction. By the final time point at 240 min, Split-Broccoli demonstrates a 124-fold increase over either Top-T (yellow) or Bottom-T (blue) alone. Mean values are shown (n = 4) with error bars to indicate standard deviations.