PCR Amplification and Transcription for Site-Specific Labeling of Large RNA Molecules by a Two-Unnatural-Base-Pair System

Figure 2

Experimental scheme of PCR amplification and transcription by a two-unnatural-base-pair system for site-specific biotin labeling of RNA molecules. DNA fragments containing the Ds-Px pair at the internal positions were prepared by a fusion PCR method. The original plasmid sequence, used as the PCR template, is shown at the top. The CDsN sequence (N = T, G, A, or C) in the P2 primer is integrated into the 86-bp DNA fragments. The 282-bp products generated by fusion PCR were used as templates for T7 transcription with Biotin-PaTP.