FIGURE 2.

Formation of the TP-dAMP initiation complex using as template ssDNA (A) and reconstituted TP-DNA (B). The assays were carried out essentially as described under “Experimental Procedures” in the presence of 1 mm MnCl₂. The position of the TP-dAMP is indicated. TP-DNA, initiation reaction using as template intact TP-DNA; C (control), initiation reaction using as template TP-DNA incubated at the same temperature and in the same reaction mixture as reconstituted DNA but in the absence of Vent polymerase and oligonucleotide; O (oligonucleotide), initiation reaction using as template the amount of oligonucleotide used in the hybridization but without TP-DNA; D (degraded), initiation reaction using as template TP-DNA with the 3′ ends degraded but not hybridized to the oligonucleotide.