FIGURE 2.

TCC assembly according to the ITR ends. A, diagram of the different types of ITR used. UC-ITR, PN-ITR, and nontransferred strand (NTS) cleaved 3 bp inside the ITR; PC-ITR, nontransferred strand, and transferred strand (TS) are cleaved. B, TCC assembly with various ITRs. For each ITR, PIC assembly was assayed using labeled ITRs (*) and Mg²⁺, at either 4 or 30 °C (as specified). TCC were assembled using cold PICs and a labeled target (*) without EDTA, at 4 °C when the PICs were preformed at 4 °C (lanes 3, 7, and 11) and with or without EDTA at 30 °C when the PICs were preformed at 30 °C (lanes 4, 8, and 12). C, for quantification, the percentage of PIC was obtained by dividing the amount of PIC by the amount of PIC + free ITR. The percentage of TCC was obtained by dividing the amount of TCC by the amount of TCC + free target. The ratio TCC/PIC was calculated for each ITR and plotted on a graph, and the values are indicated in the table.