Patent ID: 7435562

Claim:
A method of generating a double-stranded DNA molecule, the method comprising steps of: providing a template nucleic acid molecule; contacting the template nucleic acid molecule with a first primer that hybridizes thereto, the first primer including at least one termination residue consisting of a 2′-O-methyl residue; extending the first primer so that a nucleic acid strand complementary to that with which the first primer hybridizes is generated; contacting the complementary nucleic acid strand with a second primer that hybridizes thereto; extending the second primer with a DNA polymerase that does not copy the 2′-O-methyl residue, so that the extension reaction produces a product molecule containing a first overhang; providing a second double-stranded DNA molecule containing a second overhang at least partly complementary to the first overhang; and combining the second double-stranded DNA molecule with the generated double-stranded DNA molecule under conditions that allow hybridization of the first and second overhangs.