MEASURING METHOD AND MEASURING APPARATUS OF DIGITAL PCR

• Main Authors: NAKAGAWA SHIGEO, HARADA KUNIO, TANAKA JUNKO, SHIMAZAKI YUZURU
• Format: Patent
• Language: English; Japanese
• Published: 19.12.2019
• Subjects: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

To provide a novel analysis method of digital PCR.SOLUTION: A DNA detection method includes: a process of dividing DNA solution containing fluorescent-labeled probe or DNA intercalator and DNA to be detected into a plurality of fractions; a process of performing nucleic acid amplification reaction in the fractions; a process of measuring fluorescence intensity with temperature changes; a process of calculating melting temperature of DNA double-strand which is measured based on changes of the fluorescence intensity with the temperature change; and a process of calculating a ratio between the fluorescence intensity of first temperature with the temperature changes and the fluorescence intensity of second temperature lower than the first temperature.SELECTED DRAWING: Figure 8 【課題】新たなデジタルＰＣＲの解析方法を提供すること。【解決手段】蛍光標識プローブまたはＤＮＡインターカレーターと検出対象のＤＮＡを含有するＤＮＡ溶液を複数の画分に分割する工程と、前記画分の中で核酸増幅反応を行う工程と、温度変化に伴って蛍光強度を測定する工程と、前記温度変化に伴う前記蛍光強度の変化に基づいて計測されるＤＮＡ二重鎖の融解温度を算出する工程と、前記温度変化に伴う第１の温度の蛍光強度に対する、第１の温度より低い第２の温度の蛍光強度の比を算出する工程と、を含む、ＤＮＡ検出方法とする。【選択図】図８