WO2010091046A3 - Systems and methods for high throughput, high fidelity, single molecule nucleic acid sequencing using time multiplexed excitation - Google Patents

Systems and methods for high throughput, high fidelity, single molecule nucleic acid sequencing using time multiplexed excitation Download PDF

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Publication number
WO2010091046A3
WO2010091046A3 PCT/US2010/022988 US2010022988W WO2010091046A3 WO 2010091046 A3 WO2010091046 A3 WO 2010091046A3 US 2010022988 W US2010022988 W US 2010022988W WO 2010091046 A3 WO2010091046 A3 WO 2010091046A3
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WO
WIPO (PCT)
Prior art keywords
nucleic acid
illumination beam
target nucleic
high throughput
multiplexed excitation
Prior art date
Application number
PCT/US2010/022988
Other languages
French (fr)
Other versions
WO2010091046A2 (en
Inventor
Xioliang Sunney Xie
Peter A. Sims
William J. Greenleaf
Original Assignee
President & Fellows Of Harvard College
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by President & Fellows Of Harvard College filed Critical President & Fellows Of Harvard College
Publication of WO2010091046A2 publication Critical patent/WO2010091046A2/en
Publication of WO2010091046A3 publication Critical patent/WO2010091046A3/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing

Abstract

A method is disclosed of using microscopy in high throughput sequencing of a target nucleic acid. The method includes the steps of disposing in a microreactor a mixture in solution phase comprising a copy of a target nucleic acid, providing a first illumination beam having a center frequency of ωi that is selected to effect fluorescence of a first label associated with a first nucleotide, providing a second illumination beam having a center frequency of ct^ that is selected to effect fluorescence of a second label associated with a second nucleotide, and modulating the first illumination beam and the second illumination beam such that a multiplexed excitation beam is provided that includes illumination from the first illumination beam and the second illumination beam. The method also includes the steps of directing and focusing the multiplexed excitation beam toward a common focal volume, and detecting radiation at an array detector from within the common focal volume. The radiation is associated with the incorporation of an individual nucleotide during template-dependent replication of a copy of the target nucleic acid to achieve high throughput sequencing of the target nucleic acid.
PCT/US2010/022988 2009-02-03 2010-02-03 Systems and methods for high throughput, high fidelity, single molecule nucleic acid sequencing using time multiplexed excitation WO2010091046A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US14950309P 2009-02-03 2009-02-03
US61/149,503 2009-02-03

Publications (2)

Publication Number Publication Date
WO2010091046A2 WO2010091046A2 (en) 2010-08-12
WO2010091046A3 true WO2010091046A3 (en) 2011-04-07

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2010/022988 WO2010091046A2 (en) 2009-02-03 2010-02-03 Systems and methods for high throughput, high fidelity, single molecule nucleic acid sequencing using time multiplexed excitation

Country Status (1)

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WO (1) WO2010091046A2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2602063A (en) * 2020-12-17 2022-06-22 Sumitomo Chemical Co Sequencing method
WO2023230207A1 (en) * 2022-05-27 2023-11-30 President And Fellows Of Harvard College Fluorogenic nucleosides

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5784157A (en) * 1995-11-21 1998-07-21 The Research Foundation Of State University Of New York Method and apparatus for identifying fluorophores
US20030058440A1 (en) * 2001-08-28 2003-03-27 Scott Graham B. I. Pulsed-multiline excitation for color-blind fluorescence detection
US20050227231A1 (en) * 2001-10-04 2005-10-13 Dimitri Tcherkassov Device for sequencing nucleic acid molecules
US20070070349A1 (en) * 2005-09-23 2007-03-29 Helicos Biosciences Corporation Optical train and method for TIRF single molecule detection and analysis
US20070109536A1 (en) * 2003-06-20 2007-05-17 The Regents Of The University Of California Modulated excitation fluorescense analysis

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5784157A (en) * 1995-11-21 1998-07-21 The Research Foundation Of State University Of New York Method and apparatus for identifying fluorophores
US20030058440A1 (en) * 2001-08-28 2003-03-27 Scott Graham B. I. Pulsed-multiline excitation for color-blind fluorescence detection
US20050227231A1 (en) * 2001-10-04 2005-10-13 Dimitri Tcherkassov Device for sequencing nucleic acid molecules
US20070109536A1 (en) * 2003-06-20 2007-05-17 The Regents Of The University Of California Modulated excitation fluorescense analysis
US20070070349A1 (en) * 2005-09-23 2007-03-29 Helicos Biosciences Corporation Optical train and method for TIRF single molecule detection and analysis

Also Published As

Publication number Publication date
WO2010091046A2 (en) 2010-08-12

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