Fig. 2: Yield, fidelity, and NMR sample quality of SegModTeX. | Nature Communications

Fig. 2: Yield, fidelity, and NMR sample quality of SegModTeX.

From: Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA

Fig. 2

a HBV-ε seg1 (25-nt, left), tRNAlys3 seg1 (20-nt, center), and 7SK-SL1apical seg1 (41-nt, right) RNAs are fully extended by 1, 12, and 29 nucleotides, respectively. Loaded reaction equivalents with respect to Seg1 input are denoted due to higher dilution volumes being required for DNase treatment b Accurate incremental extensions of HBV-ε seg1 (25-nt, lane 1) from various NTP mix combinations (lanes 2 to 5). Predicted extensions occur exactly as delimited by complementary rNTPs, indicating a low promiscuity of the polymerase. c Lack of HBV-ε seg1 (25-nt, lane 1) extension with mismatched 3’-end base pairs (C:dC, U:dC, A:dC, lanes 3-5, respectively) indicates high fidelity. Additionally, a 26-nt HBV-ε seg1 with a G:dT mismatch extends inefficiently (lane 2), demonstrating a strong preference for canonical Watson-Crick base pairing. d 2D-NOESY overlays of 7SK-SL1apical (56-nt) constructed by T7 polymerase (black) and by SegModTeX extension of a 28-nt seg1 (red). Source data are provided as a Source Data file.

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