Figure 6.

Characterization of the in vitro integration assay. (a) The integration assay was carried out with increasing ratios of Rep to substrates (0:1, 0.5:1, 5:1, 50:1, and 500:1). (b) The integration reaction was incubated at 34°C for 24 h, aliquots were removed at eight points (0, 0.25, 0.5, 1, 2, 6, 18, and 24 h), and the reaction was stopped by adding SDS. (c) Lane 1 represents the integration assay as described. The integration reaction was carried out in absence of Mg²⁺ (lane 2), ATP, creatine phosphate, and creatine phosphokinase (lane 3), HeLa extract (lane 4), nucleotides (lane 5), or DNA substrates (lane 6). Lane M contains the DNA molecular weight marker VI from Boehringer Mannheim.