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    • 6. 发明专利
    • Quantification method of target nucleic acid
    • JP4438110B2
    • 2010-03-24
    • JP18643498
    • 1998-07-01
    • 東ソー株式会社
    • 寿一 斎藤哲也 石塚敬彦 石黒
    • C12Q1/68G01N33/50
    • C12Q1/686C12Q1/6865C12Q2563/173C12Q2527/125C12Q2521/107C12Q2521/119C12Q2563/107
    • A simple and accurate method for assay of a single-stranded RNA containing a specific nucleic acids sequence in a sample at almost constant temperature by using at least the following reagents (A) to (I), which comprises a step of adding the reagents (A) to (I) one by one (in any order), in combinations of at least two or all at once and a step of measuring a fluorescent signal in the presence of the reagent (I) at least once after addition of at least the reagents (A) to (H); (A) a first single-stranded oligonucleic acid complementary to a sequence neighboring the 5' end of the specific nucleic acids sequence in the single-stranded RNA, (B) a second single-stranded oligo DNA complementary to a 3'-end sequence within the specific nucleic acids sequence, (C) an RNA-dependent DNA polymerase, (D) deoxyribonucleoside triphosphates, (E) a third single-stranded oligo DNA having (1) a promoter sequence for a DNA-dependent RNA polymerase, (2) an enhancer sequence for the promoter and (3) a 5'-end sequence within the specific nucleic acids sequence, in this order from the 5' end, (F) a DNA-dependent DNA polymerase, (G) a DNA-dependent RNA polymerase, (H) ribonucleoside triphosphates, and (I) a fourth single-stranded oligo DNA complementary to the specific nucleic acids sequence which is labeled so that it gives off a measurable fluorescent signal on hybridization with a nucleic acid containing the specific nucleic acids sequence.