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. Author manuscript; available in PMC: 2018 Apr 7.
Published in final edited form as: Methods Enzymol. 2017 Apr 7;591:187–210. doi: 10.1016/bs.mie.2017.03.004

Figure 2.

Figure 2

Method outline to replace an OG-containing base pair with a dNaM:d5SICS or dMMO2:d5SICS unnatural DNA base pairs. (A) The dNaM:d5SICS and dMMO2:d5SICS structures. (B) Steps in the method to replace an OG-containing base pair with the dNaM:d5SICS unnatural DNA base pair. aFor clarity only dNaMTP is shown in the figure; however, this step can also be achieved by allowing the polymerase to insert d5SICSTP. bFor the sake of brevity, amplicons resulting from the unnatural nucleotide containing strand are shown and sequenced. In practice, both strands from Step IV will amplify yielding a mixture of two duplexes, one with the unnatural base pair and the other without. A method to purify the unnatural base pair-containing duplex away from the other duplex is presented in a note below.