Methods and kits for preparing and detecting RNA probes

摘要

A method of preparing labeled RNA probe by reacting RNA polymerase in the presence of a DNA fragment comprising a promoter sequence for the RNA polymerase and substrates of the RNA polymerase. In the method, at least one of said substrates comprises said label, and said RNA polymerase is mutant RNA polymerase where at least one of the amino acids of wild type RNA polymerase has been modified to permit incorporation of the substrate having a label or to improve the incorporation of the substrate having a label. A method of detecting targeted nucleic acid in which targeted nucleic acid and labeled RNA probe prepared by the above method are mixed and RNA probe that has hybridized with the targeted nucleic acid is selectively detected. A kit for preparing labeled RNA probe comprising (1) RNA polymerase, (2) DNA comprising a promoter sequence for said RNA polymerase, (3) substrates of said RNA polymerase, and (4) optionally an instruction manual. In the kit, at least one of said substrates comprises a label and said RNA polymerase is mutant RNA polymerase where at least one of the amino acids of wild type RNA polymerase has been modified to permit incorporation of said substrate having a label or to improve the incorporation of said substrate having a label.