Patent ID: 8206909

Claim:
A method of modifying a nucleic acid, comprising the steps of: contacting an Original DNA, comprising a Target DNA Fragment having a Modification Target flanked by two restriction sites, with a DNA polymerase and two Starter Primers selected for amplifying the Target DNA Fragment, such that the Starter Primers anneal to segments of the Original DNA at the ends of the Target DNA Fragment; amplifying, with the Starter Primers, a portion of the Original DNA comprising the Target DNA Fragment, generating a Starter DNA; self-ligating the Starter DNA with a DNA ligase, forming a Closed Starter DNA; contacting the Closed Starter DNA with a DNA polymerase and two Opener Primers selected for causing a modification within the Target DNA Fragment, such that the Opener Primers anneal to opposite strands of the double stranded Target DNA Fragment such that the 5′ ends of the Opener Primers are proximal and the 3′ ends are distal relative to each other; amplifying the Closed Starter DNA with the Opener Primers, generating an Intermediate DNA; and forming the modified nucleic acid by the steps of: self-ligating the Intermediate DNA with DNA ligase, forming a Closed Intermediate DNA; and, optionally, amplifying the Closed Intermediate DNA by contacting the Closed Intermediate DNA with the Starter Primers and a DNA polymerase, generating a Linear Modified DNA containing the modification.