Patent Evaluation: Direct Cloning of PCR Amplified Nucleic Acids

Summary

Novelty: A vector is described which allows the direct cloning of PCR amplified DNA containing a single dAMP residue, which is attatched to the 3′-end of the fragment. This direct cloning allows the efficient recovery of PCR generated DNA for diagnostic purposes and for construction.

Biology: Several thermostable DNA polymerases used for PCR add a single dAMP residue at the 3′-end of the DNA fragment. A vector which contains a single dT residue at the 3′-ends, complementary to the dAMP residues on the PCR fragment are revealed. The two can then be efficiently ligated together. The vector contains two Xcm I sites in the multiple cloning site. These sites can be cleaved to give a linearized vector with a single dT at each 3′-end. The use of Hph I for the same purpose is also reported.