Title:
DNA DETECTING METHOD AND APPARATUS THEREFOR
Document Type and Number:
WIPO Patent Application WO/2018/008083
Kind Code:
A1
Abstract:
The present invention has been conceived for the purpose of providing a new DNA detecting method and an apparatus therefor. In other words, provided is a DNA detecting method including: a step for manufacturing droplets by adding, to oil, a DNA solution that contains fluorescent labeling probes, DNA intercalators, and DNA molecules serving as detection targets; a step for performing PCR on the droplets; and a step for measuring fluorescence coming from the fluorescent labeling probes and fluorescence coming from the DNA intercalators, wherein the concentration of the DNA solution is such that the amount of the DNA molecules serving as the detection targets does not exceed one molecule per droplet. Additionally provided is a DNA detection apparatus provided with: a droplet manufacturing unit, a thermal cycler unit, and a fluorescence detection unit that perform the aforementioned steps, respectively.
Inventors:
TANAKA JUNKO (JP)
KOHARA YOSHINOBU (JP)
HARADA KUNIO (JP)
ISHIGE YU (JP)
KOHARA YOSHINOBU (JP)
HARADA KUNIO (JP)
ISHIGE YU (JP)
Application Number:
PCT/JP2016/069887
Publication Date:
January 11, 2018
Filing Date:
July 05, 2016
Export Citation:
Assignee:
HITACHI LTD (JP)
International Classes:
C12Q1/68; C12M1/00; C12N15/09; G16B25/20
Foreign References:
| JP2013521764A | 2013-06-13 | |||
| JP2014510538A | 2014-05-01 |
Other References:
QX200 DROPLET DIGITAL PCR & AUTODG, April 2015 (2015-04-01), pages 1 - 20, Retrieved from the Internet [retrieved on 20160928]
MCDERMOTT GEOFFREY P. ET AL.: "Multiplexed Target Detection Using DNA-Binding Dye Chemistry in Droplet Digital PCR", ANALYTICAL CHEMISTRY, vol. 85, 2013, pages 11619 - 11627, XP055152406
LIND KRISTINA ET AL.: "Combining sequence- specific probes and DNA binding dyes in real- time PCR for specific nucleic acid quantification and melting curve analysis", BIOTECHNIQUES, vol. 40, no. 3, 2006, pages 315 - 319, XP002629889
MCDERMOTT GEOFFREY P. ET AL.: "Multiplexed Target Detection Using DNA-Binding Dye Chemistry in Droplet Digital PCR", ANALYTICAL CHEMISTRY, vol. 85, 2013, pages 11619 - 11627, XP055152406
LIND KRISTINA ET AL.: "Combining sequence- specific probes and DNA binding dyes in real- time PCR for specific nucleic acid quantification and melting curve analysis", BIOTECHNIQUES, vol. 40, no. 3, 2006, pages 315 - 319, XP002629889
Attorney, Agent or Firm:
ISSHIKI & CO. (JP)
Download PDF: