Method of determining dna base sequence

摘要

A transcriptional sequence method whereby a high SN ratio can be obtained in sequence analysis with the use of capillary and longer and more accurate base sequence data can be obtained in a single reaction. More specifically, a method of determining a DNA base sequence involving the step of obtaining a nucleic acid transcription product with the use of an RNA polymerase, a template DNA having a promoter sequence for the RNA polymerase and substrates of the RNA polymerase. The substrates of the RNA polymerase involve a 3'-deoxynucleotide derivative. The RNA polymerase is a mutant RNA polymerase derived from a wild type RNA polymerase by substitution of at least one amino acid or deletion of at least one amino acid. The substitution and/or deletion of the amino acid(s) are performed so that the mutant RNA polymerase has an enhanced capability of incorporating the 3'-deoxynucleotide derivative as a substrate compared with the corresponding wild type RNA polymerase.