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. 2007 Apr 22;35(9):3118–3127. doi: 10.1093/nar/gkm168

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© 2007 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — (a) Extension of primers using the indicated diastereoisomers of dPTPαS with Taq and 9°N (modified) DNA polymerases for the times indicated. The positions of migration of the unextended primer, primer extended by 1 nt and the primer extended by 2 nt, are indicated by P(N), N + 1, N + 2, respectively. The amount of oligonucleotide loaded to mark the position where the primer runs (lane 1) was less than for other lanes in the gel; (b) Exo III digestion of the products of 2-min primer extension reactions using the indicated diastereomers (S or R) of alpha-thio-dPTP (from Figure 5a). Each primer extension product was purified with a QIAquick column and digested with 20 Units of Exo III for the indicated times. The control shows the degradation of 5′-³²P-labeled primer Z-SS-S19 (25mer) in a duplex with unlabeled Z-51-Temp, establishing that these are degraded by Exo III.