Patent ID: 6391548
Filing Date: 2002-05-21
Classification: C12N,C12Q,Y10S

Abstract:
A method of introducing a specific mutation into a selected DNA molecule for mutagenesis, wherein said DNA molecule is a double-stranded circular DNA molecule, said method comprising the steps of:annealing a first mutagenic primer and a second mutagenic primer to said DNA molecule, wherein said first mutagenic primer comprises a region that is complementary to the second mutagenic primer and wherein said first and second mutagenic primers, synthesizing by means of a linear cyclic amplification reaction a first mutagenized DNA strand comprising said first mutagenic primer, and a second mutagenized DNA strand comprising said second mutagenic primer, wherein the first mutagenized DNA strand and the second mutagenized DNA may form a double-stranded mutagenized circular DNA intermediate, and digesting said DNA molecule for mutagenesis, wherein said digestion is mediated by a selection enzyme.