Patent ID: 6333159
Filing Date: 2001-12-25
Classification: C12N,C12Q

Abstract:
A process for amplifying a specific nucleic acid sequence, said process comprising:(a) if the nucleic acid sequence is double stranded, separating the strands and denaturing intrastrand structures;(b) treating the single strands with oligonucleotide primers, under conditions such that an extension product of each primer is synthesized, using a mutated DNA polymerase comprising at least one mutation, said mutant DNA polymerase exhibiting substantially reduced polymerase activity when compared to the same polymerase without the at least one mutation at 25.degree. C. and substantially similar polymerase activity when compared to the same polymerase without the at least one mutation at 68.degree. C., where the extension product is complementary to each DNA strand;(c) separating the primer extension products from the templates on which they are synthesized to produce single-stranded molecules; and(d) repeating steps b and c at least once.