Primer-independent RNA synthesis on RNA templates with NV3Dpol. (A) The sequence of RNA templates. (B) NV3Dpol (5 pmol) was incubated with (i) Temp(GGG-CCC), (ii) Temp(GGG-GGG), (iii) Temp(GGG-CCA) or (iv) Temp(GGG-UAC) (5 pmol each), and sampled at 0, 60, 120, 180 min respectively (reaction volume = 20 μL). Each reaction products were separated on an 8 M urea denaturing 10% PAGE, and imaged with Fx imager after SYBRgreenII staining. M; 10 bp DNA step ladder marker (Promega). Black arrow indicates the mobility of each RNA templates (50 nts). For bands indicated by asterisk and short arrows, see text. Other constant bands were from components of cell-free translation system.