I'm trying to obtain linear ssDNA between 5-10 kb long. I have tried cleaving M13mp18 plasmid (circular, 7 kb) with Swa I endonuclease after annealing a 30 bp primer over the restriction site, but I have not been successful in obtaining my desired product. Does anyone know why this might be? I have already tried various optimization steps by varying enzyme concentration, incubation time, etc. Is there another enzyme that might be better?
I have begun looking into other techniques to obtain my final product, such as PCR and separating a double-stranded linear DNA, but I am unclear as to the best approach. Any help would be greatly appreciated.