Patent ID: 6242222
Filing Date: 2001-06-05
Classification: C12N

Abstract:
A method for performing programmed sequential mutagenesis of nucleic acids comprising providing an original template nucleic acid or nucleic acid analog including a first target sequence;providing a first mutagenic primer nucleic acid or nucleic acid analog including a first target-binding sequence capable of hybridizing to said first target sequence under conditions permissive for polymerase mediated nucleic acid synthesis, but which is non-complementary to said first target sequence at one or more nucleotide or nucleotide analog positions;subjecting said original template and said first mutagenic primer to a first round of nucleic acid synthesis to produce a first mutagenized nucleic acid or nucleic acid analog including a second target sequence which is neither identical nor complementary to any sequence in said original template nucleic acid or nucleic acid analog;providing a second mutagenic primer nucleic acid or nucleic acid analog including a second target-binding sequence capable of hybridizing to said second target sequence under conditions permissive for polymerase mediated nucleic acid synthesis, but which is non-complementary to said second target sequence atone or more nucleotide or nucleotide analog positions; andsubjecting said first mutagenized nucleic acid or nucleic acid analog and said second mutagenic primer to a second round of nucleic acid synthesis to produce a second mutagenized nucleic acid or nucleic acid analog including a third target sequence which is neither identical nor complementary to any sequence in said original template or first mutagenized nucleic acid or nucleic acid analogs.