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. 2011 Feb 3;17(10):2903–2915. doi: 10.1002/chem.201001952

Figure 2.

Figure 2

The use of 3′-O-(2-cyanoethyl)-dNTP derivatives for primer extension by reverse transcriptases. A) DNA substrate used (* indicates the radiolabelled 5′-terminus). The position in the template strand that directs the nucleotide to be incorporated is in bold type. B) Incorporation of 3′-O-(2-cyanoethyl)-dTTP. Lanes 1: starting-material primer, polymerases; 2 and 3: M-MuLV; 4 and 5: RevertAid M-MuLV; 6 and 7: RevertAid H minus M-MuLV; 8 and 9: AMV (cloned); 10 and 11: ThermoScript; 12 and 13: SuperScript III; 14 and 15: Transcriptor. dTTP was added into the reaction mix after the primer extension step, as indicated for lanes 3, 5, 7, 9, 11, 13, and 15. C) Time course of incorporation of 3′-O-(2-cyanoethyl)-dTTP by RevertAid M-MuLV reverse transctiptase in the presence of Mn2+ ions. D) Extension of the corresponding substrates by the dye-labelled 3′-O-(2-cyanoethyl)-modified nucleotides 2–5 during a reaction time of 5 min.