Fig. 4: Tri/tetranucleotide PureCap analogs are incorporated by T7 RNAP in a model system. | Nature Communications

Fig. 4: Tri/tetranucleotide PureCap analogs are incorporated by T7 RNAP in a model system.

From: Cap analogs with a hydrophobic photocleavable tag enable facile purification of fully capped mRNA with various cap structures

Fig. 4: Tri/tetranucleotide PureCap analogs are incorporated by T7 RNAP in a model system.The alt text for this image may have been generated using AI.

a Three types of promoter sequences were tested using 60-bp dsDNA. The estimated position of cap analogs when they were incorporated into RNA was depicted in the figure. Major or minor species were shown in blue or gray color, respectively. b, c dPAGE analysis of the reaction. An aliquot was taken from the reaction mixture and analyzed (upper panel in b, c). Or they were analyzed after being cleaved by DNAzyme 10–23 to align the heterogeneous 3′ ends of the RNA transcripts (lower panel in b). b Effect of the promoter sequence on the incorporation of the cap analogs. The concentrations of NTPs and cap analogs were 2 mM. c Analysis of IVT on the template with Type III ϕ6.5 promoter. NTP other than GTP and cap analog concentrations were set at 2 mM, and GTP concentrations were set at either 0.5 or 2 mM. Each experiment was repeated independently at least three times to obtain similar results. Source Data are provided with this paper.

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