Patent ID: 7727745

Claim:
A method of producing a single-stranded polynucleotide molecule, which comprises: providing a template polynucleotide molecule having a sequence of bases complementary to a target polynucleotide molecule, said template sequence including at least one uracil base; annealing to the template molecule an intermediate polynucleotide molecule having a sequence complementary to the template molecule; cutting the intermediate polynucleotide molecule into two fragments while annealed to said template molecule and extending one of said fragments to replace the other by nucleotide polymerization employing nucleotide monomers of increased molecular weight or ionic charge compared to nucleotide monomer residues forming said intermediate molecule, thereby forming said target polynucleotide molecule having sequence complementary to said template; and separating said template from said target polynucleotide molecule, thereby forming a single-stranded target polynucleotide molecule.