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. 2016 Jun 17;44(14):6883–6895. doi: 10.1093/nar/gkw567

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — The influence of polymerase fidelity and acceptor template sequence. (A) Representative sym/sub template switching assays using the poliovirus polymerase variants indicated. Denaturing PAGE was used to separate reaction products at 2, 4, 8, 10, 20 and 30 min after addition of the extension mix. (B) Quantification of the transfer product generated by WT (•), H273R (▴) and G64S (▪) polymerases as a percentage of total RNA products per sample over time. Results are the average from three independent experiments and error bars indicate standard deviation. (C) Denaturing PAGE gel of in vitro sym/sub reactions quenched at 30 minutes after the addition of the extension mix containing complementary (WT) or acceptor RNAs truncated at the 3′ end by one, two, or three nucleotides (D31, D32 and D33 respectively) as shown.