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. Author manuscript; available in PMC: 2019 Jan 18.
Published in final edited form as: Chembiochem. 2017 Dec 7;19(2):142–146. doi: 10.1002/cbic.201700538

Figure 3.

Figure 3

(A) Purity of recombinant T7RNAP used in this study, as demonstrated by SDS-PAGE [10% (w/v) polyacrylamide] analysis. (B) One µl from a 100-µl IVT performed using either T7RNAP WT (lane 1) or P266L (lane 2) was electrophoresed on a 2% (w/v) agarose gel and stained with ethidium bromide; samples examined were from post-DNase I treatment. Lane 3 has 1 µg of the final pre-tRNACys 55–23 RNA that was generated using the OPME (post-IVT RppH and Xrn-1) approach. (C) Fluorescence (λexc, 312 nm) of 5 µM RNA obtained after IVT with T7RNAP WT (1) or P266L (2), or using the OPME method with P266L (3). (D) RNA yields (grey) and % incorporation of thG (purple) with T7RNAP WT (1) or P266L (2), or using the OPME method with P266L (3). Mean and standard deviation values were calculated from three independent measurements.