Title:
METHOD FOR PERFORMING SINGLE-CELL ANALYSIS AND DEVICE THEREFOR
Document Type and Number:
WIPO Patent Application WO/2019/216271
Kind Code:
A1
Abstract:
The present invention provides a genome library production method in which cell lysis and genome amplification are performed using a simple operation. More particularly, the present invention provides a method that is for amplifying polynucleotides in cells and that comprises: a step for using a sample containing two or more cells or cell-like structures, and encapsulating the cells or cell-like structures into droplets, one for each droplet; a step for gelling the droplets to generate gel capsules; a step for performing lysis of the cells or cell-like structures by immersing the gel capsules in one or more types of reagents for lysis so as to cause the polynucleotides in the cells to be eluted in the gel capsules and to be kept in the gel capsules in a state where substances binding to the polynucleotides are removed; and a step for bringing the polynucleotides into contact with a reagent for amplification to amplify the polynucleotides in the gel capsules.
Inventors:
HOSOKAWA MASAHITO (JP)
TAKEYAMA HARUKO (JP)
NISHIKAWA YOHEI (JP)
KOGAWA MASATO (JP)
TAKEYAMA HARUKO (JP)
NISHIKAWA YOHEI (JP)
KOGAWA MASATO (JP)
Application Number:
PCT/JP2019/017952
Publication Date:
November 14, 2019
Filing Date:
April 26, 2019
Export Citation:
Assignee:
BITBIOME INC (JP)
International Classes:
C12N15/10; C12M1/00; C12Q1/6844
Domestic Patent References:
WO2015069798A1 | 2015-05-14 | |||
WO2018043533A1 | 2018-03-08 | |||
WO2017218486A1 | 2017-12-21 |
Foreign References:
JP2013511991A | 2013-04-11 | |||
JP2017532024W | ||||
JP2018089259A | 2018-06-14 |
Other References:
NOVAK, RICHARD ET AL.: "Single- cell multiplex gene detection and sequencing with microfluidically generated agarose emulsions", ANGEW CHEM INT ED ENGL., vol. 50, no. 2, 2011, pages 390 - 395, XP002629259, DOI: 10.1002/ANIE.201006089
ZHANG, HUIFA ET AL.: "Massively parallel single-molecule and single- cell emulsion reverse transcription polymerase chain reaction using agarose droplet microfluidics", ANAL CHEM., vol. 84, no. 8, 2012, pages 3599 - 3606, XP002745401, DOI: 10.1021/ac2033084
KUMARESAN, PALANI ET AL.: "High-throughput single copy DNA amplification and cell analysis in engineered nanoliter droplets", ANAL CHEM., vol. 80, no. 10, 2008, pages 3522 - 3529, XP055019414, DOI: 10.1021/ac800327d
NAT BIOTECHNOL, vol. 35, no. 7, July 2017 (2017-07-01), pages 640 - 646
NATURE METHODS, vol. 13, 2016, pages 759 - 762
BANKEVICH ET AL., JOURNAL OF COMPUTATIONAL BIOLOGY, vol. 19, no. 5, 2012, pages 455 - 477, Retrieved from the Internet
GUREVICH ET AL., BIOINFORMATICS, vol. 29, no. 8, 2013, pages 1072 - 5
PARKS ET AL., GENOME RESEARCH, vol. 25, 2015, pages 1043 - 1055
BOWERS ET AL., NATURE BIOTECHNOLOGY, vol. 35, no. 8, 2017, pages 725 - 731
See also references of EP 3812460A4
ZHANG, HUIFA ET AL.: "Massively parallel single-molecule and single- cell emulsion reverse transcription polymerase chain reaction using agarose droplet microfluidics", ANAL CHEM., vol. 84, no. 8, 2012, pages 3599 - 3606, XP002745401, DOI: 10.1021/ac2033084
KUMARESAN, PALANI ET AL.: "High-throughput single copy DNA amplification and cell analysis in engineered nanoliter droplets", ANAL CHEM., vol. 80, no. 10, 2008, pages 3522 - 3529, XP055019414, DOI: 10.1021/ac800327d
NAT BIOTECHNOL, vol. 35, no. 7, July 2017 (2017-07-01), pages 640 - 646
NATURE METHODS, vol. 13, 2016, pages 759 - 762
BANKEVICH ET AL., JOURNAL OF COMPUTATIONAL BIOLOGY, vol. 19, no. 5, 2012, pages 455 - 477, Retrieved from the Internet
GUREVICH ET AL., BIOINFORMATICS, vol. 29, no. 8, 2013, pages 1072 - 5
PARKS ET AL., GENOME RESEARCH, vol. 25, 2015, pages 1043 - 1055
BOWERS ET AL., NATURE BIOTECHNOLOGY, vol. 35, no. 8, 2017, pages 725 - 731
See also references of EP 3812460A4
Attorney, Agent or Firm:
YAMAMOTO Shusaku et al. (JP)
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