Patent ID: 6596494
Filing Date: 2003-07-22
Classification: C12N,C12P,C12Q

Abstract:
A method for determining sequence of a nucleic acid molecule, comprising the steps of:a) synthesizing a nucleic acid molecule from a RNAP promoter sequence in a reaction mixture containing a mutant T7-type RNA polymerase in each of four separate reactions, wherein the T7-type RNA polymerase is selected from the group consisting of T3, &phgr;I, &phgr;IIH, W31, gh1, Y and A1122, and has a reduced discrimination between canonical and non-canonical nucleoside triphosphates, each reaction comprising at least four nucleoside triphosphates, wherein at least one nucleoside triphosphate has a nucleic acid base which is complementary to each of adenine, cytidine, guanine and uracil or thymine and a sugar with either a hydroxy or a hydrogen or a fluorine at the 2â€²-position, and further comprising a ddNTP, such that each of the four separate reactions forms a plurality of reaction products of differing length, the length of said reaction products indicating the positions or the type of base corresponding to the incorporated ddNTP, and b) evaluating the reaction products so that the sequence of the template molecule may be deduced.