Patent ID: 7309570
Filing Date: 2007-12-18
Classification: C12P,C12Q

Abstract:
1. A method for synthesizing a nucleic acid molecule comprising at least one non-canonical nucleotide, comprising the steps of: a) incubating a template nucleic acid in a reaction mixture under nucleic acid synthesis conditions containing (i) a double-mutant nucleic acid polymerase, wherein said polymerase has a reduced discrimination between canonical and non-canonical nucleoside triphosphates and wherein the polymerase is a T7 RNA polymerase comprising the mutation Y639F/H784A, and (ii) at least one non-canonical nucleoside triphosphate, wherein said non-canonical nucleoside triphosphate is incorporated into the synthesized nucleic acid in place of only one canonical nucleoside triphosphate, and b) obtaining the synthesis of a nucleic acid molecule comprising at least one non-canonical nucleotide.