Figure 1

Time course of HIV RT synthesis on the ~760 nt RNA template used in the PCR-based α-complementation assay. Shown is an autoradiogram with extension of a 20 nt 5′ P-32 end-labeled DNA primer on the RNA template used for round 1 synthesis by HIV RT (see Figure 2). Full extension of the primer resulted in a 199 nt product. A DNA ladder with nt size positions is shown on the left. Concentrations of the cations and dNTPs are indicated above the lanes. Reactions were performed for (1-r) 15 s, 30 s, 1 min, 2 min, 4 min, or 8 min with Mg²⁺, Mn²⁺, and Co²⁺, and 3.25 min, 7.5 min, 15 min, 30 min, and 1 hour with Zn²⁺. A minus enzyme control (−E) is also shown. Refer to Materials and Methods for details.