USRE41780E1 - Chemical amplification based on fluid partitioning in an immiscible liquid - Google Patents
Chemical amplification based on fluid partitioning in an immiscible liquid Download PDFInfo
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- USRE41780E1 USRE41780E1 US12/118,418 US11841808A USRE41780E US RE41780 E1 USRE41780 E1 US RE41780E1 US 11841808 A US11841808 A US 11841808A US RE41780 E USRE41780 E US RE41780E
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- sample
- microdroplets
- nucleic acid
- pcr
- partitioned sections
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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
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D473/00—Heterocyclic compounds containing purine ring systems
- C07D473/26—Heterocyclic compounds containing purine ring systems with an oxygen, sulphur, or nitrogen atom directly attached in position 2 or 6, but not in both
- C07D473/32—Nitrogen atom
- C07D473/34—Nitrogen atom attached in position 6, e.g. adenine
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
Abstract
Description
where: N=number of cycles; DL,=detection limit for optical signal [moles/liter]; X=initial number of DNA molecules; V=volume containing DNA molecules [liters]; AN=Avagadro's number [6.023×1023 molecules/mole]. From Equation E1 it is clear that N, the number of cycles until detection, decreases as V, the partitioned fluid volume, decreases.
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- (1) reducing the duration of each temperature cycle—the concentration of reactants increases by enclosing them in picoliter type volumes. Since reaction kinetics depend on the concentration of the reactant, the efficiency of a microdroplet should be higher than in an ordinary vessel (such a test tube) where the reactant quantity is infinitesimal
- (2) reducing the total number of cycles—dilution of the fluorescently generated signal is largely eliminated in such a small volume, allowing much earlier detection. This effect is directly related to the number of microdroplets formed from the initial sample/reagent pool. Since PCR is an exponential process, for example, 1000 microdroplets would produce a
signal 10 cycles faster than typical processing with bulk solutions. - (3) removing interference from competing DNA templates—given the extremely small volumes involved, it is possible to isolate a single template of the target DNA in a given microdroplet. A pL microdoplet filled with a 1 pM solution, for example, will be occupied by only one molecule on average. This makes it possible to amplify only one template in mixtures containing many kinds of templates without interference. This is extremely important in processing of real world aerosol samples containing complex mixtures of DNA from many sources, and has direct application in screening of precious cDNA libraries.
Claims (51)
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
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US12/118,418 USRE41780E1 (en) | 2003-03-14 | 2008-05-09 | Chemical amplification based on fluid partitioning in an immiscible liquid |
US12/891,733 USRE43365E1 (en) | 2003-03-14 | 2010-09-27 | Apparatus for chemical amplification based on fluid partitioning in an immiscible liquid |
US13/436,693 USRE45539E1 (en) | 2003-03-14 | 2012-03-30 | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
US14/701,392 USRE46322E1 (en) | 2003-03-14 | 2015-04-30 | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
US15/421,141 USRE47080E1 (en) | 2003-03-14 | 2017-01-31 | Chemical amplification based on fluid partitioning |
US16/115,187 USRE48788E1 (en) | 2003-03-14 | 2018-08-28 | Chemical amplification based on fluid partitioning |
Applications Claiming Priority (2)
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US10/389,130 US7041481B2 (en) | 2003-03-14 | 2003-03-14 | Chemical amplification based on fluid partitioning |
US12/118,418 USRE41780E1 (en) | 2003-03-14 | 2008-05-09 | Chemical amplification based on fluid partitioning in an immiscible liquid |
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US10/389,130 Reissue US7041481B2 (en) | 2003-03-14 | 2003-03-14 | Chemical amplification based on fluid partitioning |
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US10/389,130 Continuation US7041481B2 (en) | 2003-03-14 | 2003-03-14 | Chemical amplification based on fluid partitioning |
US12/891,733 Continuation USRE43365E1 (en) | 2003-03-14 | 2010-09-27 | Apparatus for chemical amplification based on fluid partitioning in an immiscible liquid |
US12/891,733 Reissue USRE43365E1 (en) | 2003-03-14 | 2010-09-27 | Apparatus for chemical amplification based on fluid partitioning in an immiscible liquid |
Publications (1)
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USRE41780E1 true USRE41780E1 (en) | 2010-09-28 |
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Family Applications (7)
Application Number | Title | Priority Date | Filing Date |
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US10/389,130 Ceased US7041481B2 (en) | 2003-03-14 | 2003-03-14 | Chemical amplification based on fluid partitioning |
US12/118,418 Active 2024-08-17 USRE41780E1 (en) | 2003-03-14 | 2008-05-09 | Chemical amplification based on fluid partitioning in an immiscible liquid |
US12/891,733 Active 2024-08-17 USRE43365E1 (en) | 2003-03-14 | 2010-09-27 | Apparatus for chemical amplification based on fluid partitioning in an immiscible liquid |
US13/436,693 Active 2024-08-17 USRE45539E1 (en) | 2003-03-14 | 2012-03-30 | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
US14/701,392 Active 2024-08-17 USRE46322E1 (en) | 2003-03-14 | 2015-04-30 | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
US15/421,141 Active 2024-08-17 USRE47080E1 (en) | 2003-03-14 | 2017-01-31 | Chemical amplification based on fluid partitioning |
US16/115,187 Active 2024-08-17 USRE48788E1 (en) | 2003-03-14 | 2018-08-28 | Chemical amplification based on fluid partitioning |
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US10/389,130 Ceased US7041481B2 (en) | 2003-03-14 | 2003-03-14 | Chemical amplification based on fluid partitioning |
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US12/891,733 Active 2024-08-17 USRE43365E1 (en) | 2003-03-14 | 2010-09-27 | Apparatus for chemical amplification based on fluid partitioning in an immiscible liquid |
US13/436,693 Active 2024-08-17 USRE45539E1 (en) | 2003-03-14 | 2012-03-30 | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
US14/701,392 Active 2024-08-17 USRE46322E1 (en) | 2003-03-14 | 2015-04-30 | Method for chemical amplification based on fluid partitioning in an immiscible liquid |
US15/421,141 Active 2024-08-17 USRE47080E1 (en) | 2003-03-14 | 2017-01-31 | Chemical amplification based on fluid partitioning |
US16/115,187 Active 2024-08-17 USRE48788E1 (en) | 2003-03-14 | 2018-08-28 | Chemical amplification based on fluid partitioning |
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USRE48788E1 (en) | 2021-10-26 |
US20040180346A1 (en) | 2004-09-16 |
USRE47080E1 (en) | 2018-10-09 |
USRE45539E1 (en) | 2015-06-02 |
USRE46322E1 (en) | 2017-02-28 |
US7041481B2 (en) | 2006-05-09 |
USRE43365E1 (en) | 2012-05-08 |
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